Simple method of 'on-plate' growth for improved antibody screening.
Antibody phage display is a powerful biomolecular selection technology now routinely used for refining antibody diversity towards analytes of both therapeutic and diagnostic interest. Post selection, automated robotic systems can be utilised to pick, express and analyse large numbers of putative analyte-specific clones allowing the parallel screening of thousands of antibodies in less time. Most screening techniques involve a spatial addressing process whereby the selected antibodies are extracted from the cells and analysed to verify specificity. Using a simple 'on-plate' growth and screening approach, we show that antibody-expressing clones can be simultaneously cultured and analysed rapidly in antigen-coated ELISA plate wells yielding high binding signals and saving valuable selection time, while also eliminating the necessity for antibody extraction. The utilisation of the 'on-plate' technique for the screening of Fab and scFv antibodies, and a comparative analysis with commonly used antibody extraction processes, are described.
Guo
Wei Li
WL
Biomedical Diagnostics Institute and School of Biotechnology, National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland.
Leonard
Paul
P
O'Kennedy
Richard
R
D000918:Antibody Specificity / Q000276:immunology*; D001666:Binding Sites, Antibody; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments / Q000032:analysis* / Q000235:genetics / Q000276:immunology*
D007140:Immunoglobulin Fab Fragments