Abstract
A consensus peptide sequence, QSYP, appears as an artifact during the mapping of monoclonal antibodies (MAbs) using a random peptide phage display library. Phage bearing this QSYP sequence were independently selected by four different laboratories screening separate MAb preparations with the same phage library. In each case, the QSYP sequence was selected in addition to a consensus sequence specific to the MAb. Phage that displayed the QSYP sequence were not bound by the MAb of interest, but rather bound to bovine IgG derived from the FBS present in the hybridoma growth media. The implications of this finding for the interpretation of phage library screening results and possible methods for the removal of bovine IgG from MAb preparations are discussed.
Publication types
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Evaluation Study
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
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Validation Study
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / chemistry*
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Antibodies, Monoclonal / immunology
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Artifacts*
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Bacterial Outer Membrane Proteins / chemistry
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Bacterial Outer Membrane Proteins / immunology
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Cattle
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Consensus Sequence*
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Epitope Mapping / methods
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Humans
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Immunoglobulin G / chemistry*
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Immunoglobulin G / immunology
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Mice
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Molecular Sequence Data
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Peptide Fragments / chemistry
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Peptide Fragments / immunology
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Peptide Library
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Reproducibility of Results
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Sensitivity and Specificity
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Sequence Alignment / methods*
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Sequence Analysis, Protein / methods
Substances
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Antibodies, Monoclonal
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Bacterial Outer Membrane Proteins
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Immunoglobulin G
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Peptide Fragments
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Peptide Library
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major outer membrane protein, Chlamydia pneumoniae