Journal: bioRxiv

Article Title: Structural conservation of Lassa virus glycoproteins and recognition by neutralizing antibodies

doi: 10.1101/2022.09.26.509601

Figure Lengend Snippet: (A) Summary of mAb binding to GPCs by BLI (raw data in Fig. S6). The binding efficiency is based on the relative on-rate of IgG to immobilized GPC and is indicated as follows: +++, very strong binding; ++ strong binding; +, moderate binding; -, minimal to no binding. Proposed IgG occupancy per GPC is estimated based on relative R max values under the assumption the highest R max indicates full occupancy and 37.7H has a preferred occupancy of 3 Fabs per trimer . (B) mAb neutralization of pseudoviruses derived from LASV strains of diverse lineages. The dotted line indicates 50% neutralization. Data points represent the mean with error bars indicating the SEM of three technical replicates (37.7H neutralization assay comparisons shown in Fig. S7A). (C) Thermostability of LASV LIV GPC-I53-50A in complex with indicated Fabs assessed by nanoDSF. Points represent the melting temperature ( T m ) of each complex. Each melting curve is a representative of triplicate curves with melting temperatures within ±0.1°C. (D) Synthetic matriglycan binding competition microarray of StrepTagged GPC-A binding to matriglycan with and without pre-treatment with 12.1F and 19.7E IgG. GPC-I53-50A bound to matriglycan was detected using StrepMAB antibody (Fig. S7E). (E) BLI competition analysis of immobilized GPC bound to indicated IgG and then exposed to recombinant LAMP1 at a pH of 5 (Fig. S7F).

Article Snippet: All compounds were printed on NHS-ester activated glass slides (NEXTERION® Slide H, Schott Inc.) using a Scienion sciFLEXARRAYER S3 non-contact microarray equipped with a Scienion PDC80 nozzle (Scienion Inc.).

Techniques: Binding Assay, Neutralization, Derivative Assay, Nano Differential Scanning Fluorimetry, Microarray, Recombinant

Journal: bioRxiv

Article Title: Structural conservation of Lassa virus glycoproteins and recognition by neutralizing antibodies

doi: 10.1101/2022.09.26.509601

Figure Lengend Snippet: (A) BLI sensorgrams depicting immobilized GPC-I53-50A binding to S370.7 IgG in a dose-dependent manner. IgG concentrations used were 400, 200, 100, 50, 25, and 12.5 nM. K D value determined using a 1:1 binding profile and assuming partial dissociation. (B) LIV LASV pseudovirus nutralization of LASV by S370.7. The dotted line indicates 50% neutralization. Data points represent the mean with error bars indicating the SEM of three technical replicates. (C) BLI sensorgram comparing binding of S370.7 IgG to Fab to immobilized GPC. IgG and Fab were added at an equimolar concentration of 400 nM. (D) Thermostability of LIV GPC in complex with S370.7 assessed by nanoDSF. Points represent the melting temperature (T m ). Each melting curve is a representative of triplicate curves with melting temperatures within ±0.1°C. (E) Synthetic matriglycan binding microarray of StrepTagged GPC-I53-50A bound to S370.7 IgG and detected using StrepMAB antibody. (F) BLI analysis of immobilized GPC bound to S370.7 or 25.10C IgG and then exposed to recombinant LAMP-1 at a pH of 5.

Article Snippet: All compounds were printed on NHS-ester activated glass slides (NEXTERION® Slide H, Schott Inc.) using a Scienion sciFLEXARRAYER S3 non-contact microarray equipped with a Scienion PDC80 nozzle (Scienion Inc.).

Techniques: Binding Assay, Neutralization, Concentration Assay, Nano Differential Scanning Fluorimetry, Microarray, Recombinant

Journal: Nature Communications

Article Title: Cell surface glycan engineering reveals that matriglycan alone can recapitulate dystroglycan binding and function

doi: 10.1038/s41467-022-31205-7

Figure Lengend Snippet: a Chemoenzymatic synthesis scheme of defined lengths of matriglycan. b – d Microarray binding results with the matriglycan library at 100 μM utilizing b mAb IIH6 at 5 μg mL −1 , c recombinantly-tagged LG4-LG5 domains of mouse Laminin α1 (His 8 -GFP-Lama1) at 20 μg mL −1 , and d Recombinant LASV GP1 protein at 100 μg mL −1 . For b – d , measurements were taken at n = 4 technical replicates, where bars represent the mean and error bars represent SD. Source data of b – d are provided as a Source Data file.

Article Snippet: All compounds were printed on NHS-activated Nexterion® slides purchased from Schott using a Scienion sciFLEXARRAYER S3 non-contact microarray printer equipped with a Scienion PDC80 nozzle (Scienion Inc.).

Techniques: Microarray, Binding Assay, Recombinant