Screening of phage display immunoglobulin libraries by anti-M13 ELISA and whole phage PCR

Phage display libraries are now widely used in the cloning of antigen-specific antibody fragments either as Fab (light chain plus Fd portion of the heavy chain) or sFv (light and heavy chain variable domains linked via a polypeptide). This report describes two rapid methods of screening clones durin...

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Veröffentlicht in:	Nucleic acids research 1995-07, Vol.23 (13), p.2563-2564
Hauptverfasser:	Kingshury, Gillian A., Junghans, R. P
Format:	Artikel
Sprache:	eng
Schlagworte:	Bacteriophages - genetics Base Sequence DNA - isolation & purification Enzyme-Linked Immunosorbent Assay Gene Library Immunoglobulin Fab Fragments - genetics Immunoglobulin Heavy Chains - genetics Immunoglobulin Light Chains - genetics Immunoglobulins - genetics Molecular Sequence Data Polymerase Chain Reaction
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Zusammenfassung:	Phage display libraries are now widely used in the cloning of antigen-specific antibody fragments either as Fab (light chain plus Fd portion of the heavy chain) or sFv (light and heavy chain variable domains linked via a polypeptide). This report describes two rapid methods of screening clones during phage selection protocols: the first tests for antigen specificity by anti-M13 ELISA (4) and the second establishes the presence of both the heavy and light chain of Fab clones by PCR. The vector, NPC3, is derived from pComb3.
ISSN:	0305-1048 1362-4962
DOI:	10.1093/nar/23.13.2563