Journal: bioRxiv

Article Title: Nature-inspired Circular-economy Recycling (NaCRe) for Proteins: Proof of Concept

doi: 10.1101/2020.09.23.309799

Figure Lengend Snippet: (a) Photograph of mScarlet-i, and GFP solutions obtained by NaCRe from a mixture composed of glucagon, β-lactoglobulin A, and silk fibroin, purified by 6xHis tag at C-terminus, and inspected by using Invitrogen TM E-Gel TM Safe Imager TM (emission max of the blue LED = 470 nm). The image has been taken in the lab by using an iPhone Xs. (b) Image of the Coomassie stained SDS-PAGE protein gel of the purified mScarlet-i (2-3), and GFP (4-5) shown in (a). For mScarlet-i, the calibrant expressed in E. coli cells has been added to the gel (1); red-fluorescent proteins are known to produce cleaved fragments when treated at high temperature with denaturants. The GFP protein bands (4-5) are less intense with respect to mScarlet-i ones (2-3) since the GFP sample was buffer exchanged to prepare it for the second cycle of NaCRe, as described in Supporting Information i. The gel has been used to prepare the samples prior to proteomic characterization. The visible ladder used in lane M is the Biorad Precision Plus Protein TM Unstained Protein Standards.

Article Snippet: The purified and buffer-exchanged GFP was recovered by reverse spinning at 1000 rcf, 25° C, for 2 min, in Eppendorf 5424R.

Techniques: Purification, Staining, SDS Page

Journal: Vaccine

Article Title: Assessment of SARS-CoV-2 specific CD4(+) and CD8 (+) T cell responses using MHC class I and II tetramers

doi: 10.1016/j.vaccine.2021.03.008

Figure Lengend Snippet: SARS-CoV-2 peptide exchange with QuickSwitch HLA-A*02:01 Tetramer-PE molecules. 50 µg/mL Tetramerized and R-phycoerythrin coupled HLA-A*02:01 molecules were used for the peptide exchange experiment. All HLA-A*02:01 molecules were loaded with a exiting peptide and incubated with 20 µM of competing SARS-CoV-2 peptides for 4 h at room temperature (≈ 23 °C) in the presence of a kit-determined volume of Peptide Exchange Factor #1 (PEF#1). The amount of exiting peptide removed from the MHC molecule was then determined following a capture assay and plotted on the vertical axis for each tested peptide.

Article Snippet: Following a 150 μL wash with peptide exchange assay buffer #2 (MBLI: KXT-T07001 provided in the kit, must be diluted 1:10 before use) the exchanged MHC Class II molecules were stained with 25 μL of exiting peptide antibody-FITC (MBLI: KXT-T07201 provided in the kit).

Techniques: Incubation

Journal: Vaccine

Article Title: Assessment of SARS-CoV-2 specific CD4(+) and CD8 (+) T cell responses using MHC class I and II tetramers

doi: 10.1016/j.vaccine.2021.03.008

Figure Lengend Snippet: SARS-CoV-2 peptide exchange with QuickSwitch HLA-DRB1*01:01, HLA-DRB1*04:01 and HLA-DRB1*15:01 molecules. 100 µg/mL recombinant HLA-DRB1*01:01 (Blue Bars), HLA-DRB1*04:01 (Orange Bars) and HLA-DRB1*15:01 (Gray Bars) molecules were used for the peptide exchange experiment. All MHC Class II molecules were loaded with a QuickSwitch exiting peptide and incubated with 1 mM of competing SARS-CoV-2 peptides overnight (greater than10 h) at 37 °C. The percentage of exiting peptide removed from the MHC molecule was then determined following a capture assay and plotted on the vertical axis for each tested peptide. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Following a 150 μL wash with peptide exchange assay buffer #2 (MBLI: KXT-T07001 provided in the kit, must be diluted 1:10 before use) the exchanged MHC Class II molecules were stained with 25 μL of exiting peptide antibody-FITC (MBLI: KXT-T07201 provided in the kit).

Techniques: Recombinant, Incubation