[The impact of the antibody 2F5 biotinylation on the selection of the peptides from combinatorial phage library].
The impact of monoclonal antibodies (mAb) biotinylation on the output and the repertoire of selected peptides in the biopanning procedure were tested. A comparative analysis of the peptides selected from phage library using the biotinylated and non-biotinylated mAb 2F5 was performed. It was shown that the output of peptides homologous to the native epitope was 1.7-fold higher for biotinylated antibodies, whereas the binding capacity of the selected phages with mAb 2F5 in ELISA was higher in the case of using non-biotinylated antibodies. It should be noted that the phages exposing peptides, which have 4-5 amino acid sequence similarity with the native epitope, demonstrate the highest binding affinity. The phages that expose peptides with 3 amino acid sequence similarity demonstrate different binding affinity: from the smallest to the largest. Based on the obtained data, it is safe to suggest that the rational biopanning may proceed in accordance with the task.
N S
NS
Shcherbakova
A N
AN
Chikaev
L I
LI
Karpenko
A A
AA
Il'ichev
D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics; D019921:Biotinylation; D000080908:Broadly Neutralizing Antibodies; D000939:Epitopes / Q000235:genetics; D015870:Gene Expression; D015483:HIV Antibodies; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000302:isolation & purification*; D017386:Sequence Homology, Amino Acid
C558560:2F5 monoclonal antibody; D000911:Antibodies, Monoclonal; D000080908:Broadly Neutralizing Antibodies; D000939:Epitopes; D015483:HIV Antibodies; D019151:Peptide Library; D010455:Peptides