A high throughput ultra performance size exclusion chromatography assay for the analysis of aggregates and fragments of monoclonal antibodies

2014 ◽  
Vol 2 (2) ◽  
pp. 141-156 ◽  
Author(s):  
Xiaodun Mou ◽  
Xiaoyu Yang ◽  
Hong Li ◽  
Alexandre Ambrogelly ◽  
David J Pollard
Keyword(s):  
Monoclonal Antibodies ◽  
Size Exclusion Chromatography ◽  
High Throughput ◽  
Size Exclusion ◽  
Exclusion Chromatography

scholarly journals Importance of vial shape and type on the reproducibility of size exclusion chromatography measurement of monoclonal antibodies

2016 ◽  
Vol 1032 ◽  
pp. 131-138 ◽  
Author(s):  
Marta Rodriguez-Aller ◽  
Alessandra Cusumano ◽  
Beck Alain ◽  
Davy Guillarme ◽  
Szabolcs Fekete
Keyword(s):  
Monoclonal Antibodies ◽  
Size Exclusion Chromatography ◽  
Size Exclusion ◽  
Exclusion Chromatography

scholarly journals Peptide barcoding for one-pot evaluation of sequence–function relationships of nanobodies

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yusei Matsuzaki ◽  
Wataru Aoki ◽  
Takumi Miyazaki ◽  
Shunsuke Aburaya ◽  
Yuta Ohtani ◽  
...  
Keyword(s):  
Binding Affinity ◽  
Size Exclusion Chromatography ◽  
High Throughput ◽  
Size Exclusion ◽  
Targeted Proteomics ◽  
One Pot ◽  
Unique Peptide ◽  
Exclusion Chromatography ◽  
Protein Binders

AbstractOptimisation of protein binders relies on laborious screening processes. Investigation of sequence–function relationships of protein binders is particularly slow, since mutants are purified and evaluated individually. Here we developed peptide barcoding, a high-throughput approach for accurate investigation of sequence–function relationships of hundreds of protein binders at once. Our approach is based on combining the generation of a mutagenised nanobody library fused with unique peptide barcodes, the formation of nanobody–antigen complexes at different ratios, their fine fractionation by size-exclusion chromatography and quantification of peptide barcodes by targeted proteomics. Applying peptide barcoding to an anti-GFP nanobody as a model, we successfully identified residues important for the binding affinity of anti-GFP nanobody at once. Peptide barcoding discriminated subtle changes in KD at the order of nM to sub-nM. Therefore, peptide barcoding is a powerful tool for engineering protein binders, enabling reliable one-pot evaluation of sequence–function relationships.

scholarly journals Fluorophore Absorption Size Exclusion Chromatography (FA-SEC): An Alternative Method for High-Throughput Detergent Screening of Membrane Proteins

PLoS ONE ◽  
2016 ◽  
Vol 11 (6) ◽  
pp. e0157923 ◽  
Author(s):  
Sung-Yao Lin ◽  
Xing-Han Sun ◽  
Yu-Hsuan Hsiao ◽  
Shao-En Chang ◽  
Guan-Syun Li ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Size Exclusion Chromatography ◽  
High Throughput ◽  
Size Exclusion ◽  
Alternative Method ◽  
Exclusion Chromatography
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