transglycit remodeling  (Genovis Inc)


Bioz Verified Symbol Genovis Inc is a verified supplier
Bioz Manufacturer Symbol Genovis Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Genovis Inc transglycit remodeling
    Transglycit Remodeling, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transglycit remodeling/product/Genovis Inc
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transglycit remodeling - by Bioz Stars, 2024-07
    96/100 stars

    Images

    transglycit remodeling  (Genovis Inc)


    Bioz Verified Symbol Genovis Inc is a verified supplier
    Bioz Manufacturer Symbol Genovis Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Genovis Inc transglycit remodeling
    Transglycit Remodeling, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transglycit remodeling/product/Genovis Inc
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transglycit remodeling - by Bioz Stars, 2024-07
    96/100 stars

    Images

    transglycit kit  (Genovis Inc)


    Bioz Verified Symbol Genovis Inc is a verified supplier
    Bioz Manufacturer Symbol Genovis Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Genovis Inc transglycit kit
    Transglycit Kit, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transglycit kit/product/Genovis Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transglycit kit - by Bioz Stars, 2024-07
    86/100 stars

    Images

    transglycit remodeling  (Genovis Inc)


    Bioz Verified Symbol Genovis Inc is a verified supplier
    Bioz Manufacturer Symbol Genovis Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96

    Structured Review

    Genovis Inc transglycit remodeling
    Transglycit Remodeling, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transglycit remodeling/product/Genovis Inc
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transglycit remodeling - by Bioz Stars, 2024-07
    96/100 stars

    Images

    trastuzumab with transglycit  (Genovis Inc)


    Bioz Verified Symbol Genovis Inc is a verified supplier
    Bioz Manufacturer Symbol Genovis Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Genovis Inc trastuzumab with transglycit
    CEX–UV chromatograms of intact <t>trastuzumab</t> (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.
    Trastuzumab With Transglycit, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trastuzumab with transglycit/product/Genovis Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trastuzumab with transglycit - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry"

    Article Title: Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry

    Journal: Analytical Chemistry

    doi: 10.1021/acs.analchem.2c03163

    CEX–UV chromatograms of intact trastuzumab (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.
    Figure Legend Snippet: CEX–UV chromatograms of intact trastuzumab (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.

    Techniques Used:

    CEX–CIU for mAb analysis. (a) CEX–UV chromatogram of intact trastuzumab showing the increase in CV to obtain the CIU fingerprint. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 0.2 min. (b) CEX–CIU fingerprint of the 27+ charge state of the trastuzumab peak indicated in the chromatogram. In total, four measurements were required to obtain a fingerprint from 0 to 150 V. The conformational states are labeled with numbers from 0 to 3. The replicate RMSD was 4.2%, indicating good repeatability.
    Figure Legend Snippet: CEX–CIU for mAb analysis. (a) CEX–UV chromatogram of intact trastuzumab showing the increase in CV to obtain the CIU fingerprint. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 0.2 min. (b) CEX–CIU fingerprint of the 27+ charge state of the trastuzumab peak indicated in the chromatogram. In total, four measurements were required to obtain a fingerprint from 0 to 150 V. The conformational states are labeled with numbers from 0 to 3. The replicate RMSD was 4.2%, indicating good repeatability.

    Techniques Used: Labeling

    CEX–CIU for multiplex analysis of different mAbs, including eculizumab (IgG2/4), pembrolizumab (IgG4), and trastuzumab (IgG1). (a) CEX–UV chromatogram of a mixture showing the selected peaks for CIU analysis. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 20 s. (b) Mass spectra of the selected CEX peaks, including the experimental mass of the most abundant species. (c) CEX–CIU fingerprints of the mAbs of the 27+ charge state. CEX–CIU fingerprints of the 26+ and 28+ charge states can be found in Figures S7 and S8 , respectively. (d) CIU50 values of the different detected transitions within the CIU fingerprints. (e) Linear determinant analysis of the three mAb CIU fingerprints. (f) UFS plot highlighting the critical CVs for mAb subclass identification.
    Figure Legend Snippet: CEX–CIU for multiplex analysis of different mAbs, including eculizumab (IgG2/4), pembrolizumab (IgG4), and trastuzumab (IgG1). (a) CEX–UV chromatogram of a mixture showing the selected peaks for CIU analysis. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 20 s. (b) Mass spectra of the selected CEX peaks, including the experimental mass of the most abundant species. (c) CEX–CIU fingerprints of the mAbs of the 27+ charge state. CEX–CIU fingerprints of the 26+ and 28+ charge states can be found in Figures S7 and S8 , respectively. (d) CIU50 values of the different detected transitions within the CIU fingerprints. (e) Linear determinant analysis of the three mAb CIU fingerprints. (f) UFS plot highlighting the critical CVs for mAb subclass identification.

    Techniques Used: Multiplex Assay

    Modification of the glycan moiety of trastuzumab. (a) Schematic overview of the glycan remodeling workflow. The deglycosylation was performed with or without the presence of a fucosidase, resulting in different starting points for the transglycosylation (either GlcNAcFuc or GlcNAc). (b) CEX–UV separation of glycoengineered trastuzumab at the middle level highlighting the retention time shift of the Fc domains. Corresponding mass spectra of the different Fc subunits including the (major) experimental mass are presented. Mass spectra of the F(ab′) 2 domain can be found in Figure S10 .
    Figure Legend Snippet: Modification of the glycan moiety of trastuzumab. (a) Schematic overview of the glycan remodeling workflow. The deglycosylation was performed with or without the presence of a fucosidase, resulting in different starting points for the transglycosylation (either GlcNAcFuc or GlcNAc). (b) CEX–UV separation of glycoengineered trastuzumab at the middle level highlighting the retention time shift of the Fc domains. Corresponding mass spectra of the different Fc subunits including the (major) experimental mass are presented. Mass spectra of the F(ab′) 2 domain can be found in Figure S10 .

    Techniques Used: Modification

    CEX–CIU experiments of middle-level trastuzumab samples subjected to glycan remodeling. (a) CIU fingerprints of the 14+ charge state of the Fc fragments carrying different glycans, including the initial (T0, green), deglycosylated (T1, purple), and end (G2S2, pink and G2S2F, blue) products. Differential plots including RMSD values can be found in Figure S11 . (b) ATDs extracted at 110 V evidencing the higher relative intensity of the most unfolded conformation in the cases of G2S2 and G2S2F showing greater resistance to unfolding events. (c) CIU50 analysis of the samples revealing an intermediate transition only observed in the case of sialylated variants. (d) ATDs extracted at 40 V, showing that differences in the ATD profile can also be detected in the low-energy range of the CIU fingerprints. The F(ab′) 2 CIU fingerprints showed no differences in resistance to gas-phase unfolding ( Figure S12 ).
    Figure Legend Snippet: CEX–CIU experiments of middle-level trastuzumab samples subjected to glycan remodeling. (a) CIU fingerprints of the 14+ charge state of the Fc fragments carrying different glycans, including the initial (T0, green), deglycosylated (T1, purple), and end (G2S2, pink and G2S2F, blue) products. Differential plots including RMSD values can be found in Figure S11 . (b) ATDs extracted at 110 V evidencing the higher relative intensity of the most unfolded conformation in the cases of G2S2 and G2S2F showing greater resistance to unfolding events. (c) CIU50 analysis of the samples revealing an intermediate transition only observed in the case of sialylated variants. (d) ATDs extracted at 40 V, showing that differences in the ATD profile can also be detected in the low-energy range of the CIU fingerprints. The F(ab′) 2 CIU fingerprints showed no differences in resistance to gas-phase unfolding ( Figure S12 ).

    Techniques Used:

    trastuzumab with transglycit  (Genovis Inc)


    Bioz Verified Symbol Genovis Inc is a verified supplier
    Bioz Manufacturer Symbol Genovis Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Genovis Inc trastuzumab with transglycit
    CEX–UV chromatograms of intact <t>trastuzumab</t> (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.
    Trastuzumab With Transglycit, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trastuzumab with transglycit/product/Genovis Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trastuzumab with transglycit - by Bioz Stars, 2024-07
    86/100 stars

    Images

    1) Product Images from "Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry"

    Article Title: Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry

    Journal: Analytical Chemistry

    doi: 10.1021/acs.analchem.2c03163

    CEX–UV chromatograms of intact trastuzumab (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.
    Figure Legend Snippet: CEX–UV chromatograms of intact trastuzumab (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.

    Techniques Used:

    CEX–CIU for mAb analysis. (a) CEX–UV chromatogram of intact trastuzumab showing the increase in CV to obtain the CIU fingerprint. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 0.2 min. (b) CEX–CIU fingerprint of the 27+ charge state of the trastuzumab peak indicated in the chromatogram. In total, four measurements were required to obtain a fingerprint from 0 to 150 V. The conformational states are labeled with numbers from 0 to 3. The replicate RMSD was 4.2%, indicating good repeatability.
    Figure Legend Snippet: CEX–CIU for mAb analysis. (a) CEX–UV chromatogram of intact trastuzumab showing the increase in CV to obtain the CIU fingerprint. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 0.2 min. (b) CEX–CIU fingerprint of the 27+ charge state of the trastuzumab peak indicated in the chromatogram. In total, four measurements were required to obtain a fingerprint from 0 to 150 V. The conformational states are labeled with numbers from 0 to 3. The replicate RMSD was 4.2%, indicating good repeatability.

    Techniques Used: Labeling

    CEX–CIU for multiplex analysis of different mAbs, including eculizumab (IgG2/4), pembrolizumab (IgG4), and trastuzumab (IgG1). (a) CEX–UV chromatogram of a mixture showing the selected peaks for CIU analysis. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 20 s. (b) Mass spectra of the selected CEX peaks, including the experimental mass of the most abundant species. (c) CEX–CIU fingerprints of the mAbs of the 27+ charge state. CEX–CIU fingerprints of the 26+ and 28+ charge states can be found in Figures S7 and S8 , respectively. (d) CIU50 values of the different detected transitions within the CIU fingerprints. (e) Linear determinant analysis of the three mAb CIU fingerprints. (f) UFS plot highlighting the critical CVs for mAb subclass identification.
    Figure Legend Snippet: CEX–CIU for multiplex analysis of different mAbs, including eculizumab (IgG2/4), pembrolizumab (IgG4), and trastuzumab (IgG1). (a) CEX–UV chromatogram of a mixture showing the selected peaks for CIU analysis. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 20 s. (b) Mass spectra of the selected CEX peaks, including the experimental mass of the most abundant species. (c) CEX–CIU fingerprints of the mAbs of the 27+ charge state. CEX–CIU fingerprints of the 26+ and 28+ charge states can be found in Figures S7 and S8 , respectively. (d) CIU50 values of the different detected transitions within the CIU fingerprints. (e) Linear determinant analysis of the three mAb CIU fingerprints. (f) UFS plot highlighting the critical CVs for mAb subclass identification.

    Techniques Used: Multiplex Assay

    Modification of the glycan moiety of trastuzumab. (a) Schematic overview of the glycan remodeling workflow. The deglycosylation was performed with or without the presence of a fucosidase, resulting in different starting points for the transglycosylation (either GlcNAcFuc or GlcNAc). (b) CEX–UV separation of glycoengineered trastuzumab at the middle level highlighting the retention time shift of the Fc domains. Corresponding mass spectra of the different Fc subunits including the (major) experimental mass are presented. Mass spectra of the F(ab′) 2 domain can be found in Figure S10 .
    Figure Legend Snippet: Modification of the glycan moiety of trastuzumab. (a) Schematic overview of the glycan remodeling workflow. The deglycosylation was performed with or without the presence of a fucosidase, resulting in different starting points for the transglycosylation (either GlcNAcFuc or GlcNAc). (b) CEX–UV separation of glycoengineered trastuzumab at the middle level highlighting the retention time shift of the Fc domains. Corresponding mass spectra of the different Fc subunits including the (major) experimental mass are presented. Mass spectra of the F(ab′) 2 domain can be found in Figure S10 .

    Techniques Used: Modification

    CEX–CIU experiments of middle-level trastuzumab samples subjected to glycan remodeling. (a) CIU fingerprints of the 14+ charge state of the Fc fragments carrying different glycans, including the initial (T0, green), deglycosylated (T1, purple), and end (G2S2, pink and G2S2F, blue) products. Differential plots including RMSD values can be found in Figure S11 . (b) ATDs extracted at 110 V evidencing the higher relative intensity of the most unfolded conformation in the cases of G2S2 and G2S2F showing greater resistance to unfolding events. (c) CIU50 analysis of the samples revealing an intermediate transition only observed in the case of sialylated variants. (d) ATDs extracted at 40 V, showing that differences in the ATD profile can also be detected in the low-energy range of the CIU fingerprints. The F(ab′) 2 CIU fingerprints showed no differences in resistance to gas-phase unfolding ( Figure S12 ).
    Figure Legend Snippet: CEX–CIU experiments of middle-level trastuzumab samples subjected to glycan remodeling. (a) CIU fingerprints of the 14+ charge state of the Fc fragments carrying different glycans, including the initial (T0, green), deglycosylated (T1, purple), and end (G2S2, pink and G2S2F, blue) products. Differential plots including RMSD values can be found in Figure S11 . (b) ATDs extracted at 110 V evidencing the higher relative intensity of the most unfolded conformation in the cases of G2S2 and G2S2F showing greater resistance to unfolding events. (c) CIU50 analysis of the samples revealing an intermediate transition only observed in the case of sialylated variants. (d) ATDs extracted at 40 V, showing that differences in the ATD profile can also be detected in the low-energy range of the CIU fingerprints. The F(ab′) 2 CIU fingerprints showed no differences in resistance to gas-phase unfolding ( Figure S12 ).

    Techniques Used:

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 96
    Genovis Inc transglycit remodeling
    Transglycit Remodeling, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transglycit remodeling/product/Genovis Inc
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transglycit remodeling - by Bioz Stars, 2024-07
    96/100 stars
      Buy from Supplier

    86
    Genovis Inc transglycit kit
    Transglycit Kit, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/transglycit kit/product/Genovis Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    transglycit kit - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    86
    Genovis Inc trastuzumab with transglycit
    CEX–UV chromatograms of intact <t>trastuzumab</t> (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.
    Trastuzumab With Transglycit, supplied by Genovis Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/trastuzumab with transglycit/product/Genovis Inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trastuzumab with transglycit - by Bioz Stars, 2024-07
    86/100 stars
      Buy from Supplier

    Image Search Results


    CEX–UV chromatograms of intact trastuzumab (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.

    Journal: Analytical Chemistry

    Article Title: Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry

    doi: 10.1021/acs.analchem.2c03163

    Figure Lengend Snippet: CEX–UV chromatograms of intact trastuzumab (a) and middle-level analysis of trastuzumab (b). For intact trastuzumab, acidic (deamidated proteoforms) and basic species (IsoAsp variants) were separated. The middle-level analysis resulted in two main peaks corresponding to the Fc and F(ab′) 2 fragments. Similar charge variants as for intact trastuzumab were observed on the F(ab′) 2 part.

    Article Snippet: Glycan remodeling was performed by treatment of trastuzumab with TransGLYCIT (Genovis, Lund, Sweden) according to the producer specifications with and without the presence of a fucosidase.

    Techniques:

    CEX–CIU for mAb analysis. (a) CEX–UV chromatogram of intact trastuzumab showing the increase in CV to obtain the CIU fingerprint. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 0.2 min. (b) CEX–CIU fingerprint of the 27+ charge state of the trastuzumab peak indicated in the chromatogram. In total, four measurements were required to obtain a fingerprint from 0 to 150 V. The conformational states are labeled with numbers from 0 to 3. The replicate RMSD was 4.2%, indicating good repeatability.

    Journal: Analytical Chemistry

    Article Title: Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry

    doi: 10.1021/acs.analchem.2c03163

    Figure Lengend Snippet: CEX–CIU for mAb analysis. (a) CEX–UV chromatogram of intact trastuzumab showing the increase in CV to obtain the CIU fingerprint. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 0.2 min. (b) CEX–CIU fingerprint of the 27+ charge state of the trastuzumab peak indicated in the chromatogram. In total, four measurements were required to obtain a fingerprint from 0 to 150 V. The conformational states are labeled with numbers from 0 to 3. The replicate RMSD was 4.2%, indicating good repeatability.

    Article Snippet: Glycan remodeling was performed by treatment of trastuzumab with TransGLYCIT (Genovis, Lund, Sweden) according to the producer specifications with and without the presence of a fucosidase.

    Techniques: Labeling

    CEX–CIU for multiplex analysis of different mAbs, including eculizumab (IgG2/4), pembrolizumab (IgG4), and trastuzumab (IgG1). (a) CEX–UV chromatogram of a mixture showing the selected peaks for CIU analysis. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 20 s. (b) Mass spectra of the selected CEX peaks, including the experimental mass of the most abundant species. (c) CEX–CIU fingerprints of the mAbs of the 27+ charge state. CEX–CIU fingerprints of the 26+ and 28+ charge states can be found in Figures S7 and S8 , respectively. (d) CIU50 values of the different detected transitions within the CIU fingerprints. (e) Linear determinant analysis of the three mAb CIU fingerprints. (f) UFS plot highlighting the critical CVs for mAb subclass identification.

    Journal: Analytical Chemistry

    Article Title: Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry

    doi: 10.1021/acs.analchem.2c03163

    Figure Lengend Snippet: CEX–CIU for multiplex analysis of different mAbs, including eculizumab (IgG2/4), pembrolizumab (IgG4), and trastuzumab (IgG1). (a) CEX–UV chromatogram of a mixture showing the selected peaks for CIU analysis. During elution of the selected species, the CV is increased in four steps, where each CV is maintained for 20 s. (b) Mass spectra of the selected CEX peaks, including the experimental mass of the most abundant species. (c) CEX–CIU fingerprints of the mAbs of the 27+ charge state. CEX–CIU fingerprints of the 26+ and 28+ charge states can be found in Figures S7 and S8 , respectively. (d) CIU50 values of the different detected transitions within the CIU fingerprints. (e) Linear determinant analysis of the three mAb CIU fingerprints. (f) UFS plot highlighting the critical CVs for mAb subclass identification.

    Article Snippet: Glycan remodeling was performed by treatment of trastuzumab with TransGLYCIT (Genovis, Lund, Sweden) according to the producer specifications with and without the presence of a fucosidase.

    Techniques: Multiplex Assay

    Modification of the glycan moiety of trastuzumab. (a) Schematic overview of the glycan remodeling workflow. The deglycosylation was performed with or without the presence of a fucosidase, resulting in different starting points for the transglycosylation (either GlcNAcFuc or GlcNAc). (b) CEX–UV separation of glycoengineered trastuzumab at the middle level highlighting the retention time shift of the Fc domains. Corresponding mass spectra of the different Fc subunits including the (major) experimental mass are presented. Mass spectra of the F(ab′) 2 domain can be found in Figure S10 .

    Journal: Analytical Chemistry

    Article Title: Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry

    doi: 10.1021/acs.analchem.2c03163

    Figure Lengend Snippet: Modification of the glycan moiety of trastuzumab. (a) Schematic overview of the glycan remodeling workflow. The deglycosylation was performed with or without the presence of a fucosidase, resulting in different starting points for the transglycosylation (either GlcNAcFuc or GlcNAc). (b) CEX–UV separation of glycoengineered trastuzumab at the middle level highlighting the retention time shift of the Fc domains. Corresponding mass spectra of the different Fc subunits including the (major) experimental mass are presented. Mass spectra of the F(ab′) 2 domain can be found in Figure S10 .

    Article Snippet: Glycan remodeling was performed by treatment of trastuzumab with TransGLYCIT (Genovis, Lund, Sweden) according to the producer specifications with and without the presence of a fucosidase.

    Techniques: Modification

    CEX–CIU experiments of middle-level trastuzumab samples subjected to glycan remodeling. (a) CIU fingerprints of the 14+ charge state of the Fc fragments carrying different glycans, including the initial (T0, green), deglycosylated (T1, purple), and end (G2S2, pink and G2S2F, blue) products. Differential plots including RMSD values can be found in Figure S11 . (b) ATDs extracted at 110 V evidencing the higher relative intensity of the most unfolded conformation in the cases of G2S2 and G2S2F showing greater resistance to unfolding events. (c) CIU50 analysis of the samples revealing an intermediate transition only observed in the case of sialylated variants. (d) ATDs extracted at 40 V, showing that differences in the ATD profile can also be detected in the low-energy range of the CIU fingerprints. The F(ab′) 2 CIU fingerprints showed no differences in resistance to gas-phase unfolding ( Figure S12 ).

    Journal: Analytical Chemistry

    Article Title: Online Collision-Induced Unfolding of Therapeutic Monoclonal Antibody Glyco-Variants through Direct Hyphenation of Cation Exchange Chromatography with Native Ion Mobility–Mass Spectrometry

    doi: 10.1021/acs.analchem.2c03163

    Figure Lengend Snippet: CEX–CIU experiments of middle-level trastuzumab samples subjected to glycan remodeling. (a) CIU fingerprints of the 14+ charge state of the Fc fragments carrying different glycans, including the initial (T0, green), deglycosylated (T1, purple), and end (G2S2, pink and G2S2F, blue) products. Differential plots including RMSD values can be found in Figure S11 . (b) ATDs extracted at 110 V evidencing the higher relative intensity of the most unfolded conformation in the cases of G2S2 and G2S2F showing greater resistance to unfolding events. (c) CIU50 analysis of the samples revealing an intermediate transition only observed in the case of sialylated variants. (d) ATDs extracted at 40 V, showing that differences in the ATD profile can also be detected in the low-energy range of the CIU fingerprints. The F(ab′) 2 CIU fingerprints showed no differences in resistance to gas-phase unfolding ( Figure S12 ).

    Article Snippet: Glycan remodeling was performed by treatment of trastuzumab with TransGLYCIT (Genovis, Lund, Sweden) according to the producer specifications with and without the presence of a fucosidase.

    Techniques: