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Antibody arrays for high-throughput screening of antibody-antigen interactions.
- Source :
-
Nature biotechnology [Nat Biotechnol] 2000 Sep; Vol. 18 (9), pp. 989-94. - Publication Year :
- 2000
-
Abstract
- 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.
- Subjects :
- Amino Acid Sequence
Bacteria chemistry
Bacteria genetics
Biochemistry methods
Blotting, Western
Enzyme-Linked Immunosorbent Assay
HeLa Cells
Humans
Molecular Probe Techniques
Molecular Sequence Data
Peptide Library
Protein Conformation
Proteins metabolism
Recombinant Proteins chemistry
Robotics
Serum Albumin chemistry
Serum Albumin, Bovine chemistry
Antibodies chemistry
Antigen-Antibody Reactions
Biosensing Techniques methods
Oligonucleotide Array Sequence Analysis
Proteins chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1087-0156
- Volume :
- 18
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Nature biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 10973222
- Full Text :
- https://doi.org/10.1038/79494