Antibody arrays for high-throughput screening of antibody-antigen interactions.
We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins.
de Wildt
R M
RM
MRC Laboratory of Molecular Biology and MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK. rdw@mrc-lmb.cam.ac.uk
Mundy
C R
CR
Gorick
B D
BD
Tomlinson
I M
IM
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry*; D000937:Antigen-Antibody Reactions; D001419:Bacteria / Q000737:chemistry / Q000235:genetics; D001671:Biochemistry / Q000379:methods; D015374:Biosensing Techniques / Q000379:methods*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006367:HeLa Cells; D006801:Humans; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D019151:Peptide Library; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D012371:Robotics; D012709:Serum Albumin / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry
D000906:Antibodies; D019151:Peptide Library; D011506:Proteins; D011994:Recombinant Proteins; D012709:Serum Albumin; D012710:Serum Albumin, Bovine