Selection of functional human immunoglobulin light chains from a phage-display library.
Human kappa-light chains (L chains) were amplified by the reverse transcriptase-polymerase chain reaction (PCR) and cloned into a phagemid vector. Phage particles displaying L chains were fractionated on immobilized vasoactive intestinal peptide (VIP). The resultant phage preparation displayed saturable binding of (tyr10-125I)VIP. One of the L-chain clones (hk13) was deduced to be related to subgroup I of kappa-light chains based on its nucleotide sequence. The VIP binding activity of the soluble and phage-displayed form of this L chain was confirmed by radioimmunoassay and ELISA, respectively. These observations demonstrate the potential of selecting antigen-specific L chains from phage-display libraries.
Tyutyulkova
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Department of Anesthesiology, University of Nebraska Medical Center, Omaha.
Paul
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D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D015723:Gene Library; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics* / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D013601:T-Lymphocytes / Q000276:immunology; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
D018076:DNA, Complementary; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D014660:Vasoactive Intestinal Peptide