Problems in phage-displayed Ab library

Tze Ming Leung sfnicx at hkstar.com
Fri Sep 13 06:33:33 EST 1996


Dear All,
	I have generated a phage-displayed antibody library from a hybridoma
which produces a strong Ab specific for a  15-mer peptide (P6). I call
it a 'library' because 5' degenerate primers are used in the PCR to make
VH and VL genes. This means that ScFv-genes derived from the hybridoma
contains the same VH and VL genes but with different and short leader
sequences. After four rounds of panning, I found that 90% of the
P6-binding phage (O.D. 490nm > 0.9) did not contain ScFv insert whereas
the weak binders (O.D. 490 nm < 0.2) did. I wonder why there was no good
binder with the ScFv insert. Does anybody have the same problem? What is
the solution to get rid of these unspecific binders that carry no ScFv
insert?



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