By-passing selection: direct screening for antibody-antigen interactions using protein arrays.
We have developed a system to identify highly specific antibody-antigen interactions by protein array screening. This removes the need for selection using animal immunisation or in vitro techniques such as phage or ribosome display. We screened an array of 27 648 human foetal brain proteins with 12 well-expressed antibody fragments that had not previously been exposed to any antigen. Four highly specific antibody-antigen pairs were identified, including three antibodies that bind proteins of unknown function. The target proteins were expressed at a very low copy number on the array, emphasising the unbiased nature of the screen. The specificity and sensitivity of binding demonstrates that this 'naive' screening approach could be applied to the high throughput isolation of specific antibodies against many different targets in the human proteome.
Holt
L J
LJ
MRC Laboratory of Molecular Biology and MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.
Büssow
K
K
Walter
G
G
Tomlinson
I M
IM
D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D015153:Blotting, Western; D018076:DNA, Complementary; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000276:immunology*; D019151:Peptide Library; D020543:Proteome; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000906:Antibodies; D018076:DNA, Complementary; D007128:Immunoglobulin Fragments; D009419:Nerve Tissue Proteins; D019151:Peptide Library; D020543:Proteome; D011994:Recombinant Proteins