Development of naïve phage display VHH libraries from Indian camel

L DASH; S SUBRAMANIAM; S A KHULAPE; B R PRUSTY; K PARGAI; S D NARNAWARE; N V PATIL; B PATTNAIK

doi:10.56093/ijans.v86i8.60596

Authors

L DASH ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India
S SUBRAMANIAM ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India
S A KHULAPE ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India
B R PRUSTY ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India
K PARGAI ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India
S D NARNAWARE ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India
N V PATIL ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India
B PATTNAIK ICAR-Project Directorate on Foot and Mouth Disease, Mukteswar, Uttarakhand 263 138 India

https://doi.org/10.56093/ijans.v86i8.60596

Keywords:

Nanobody, Phagemid, Redundancy, Single pot library

Abstract

The present study describes the construction of naïve camelid VHH single pot library from lymphocytes of non- immunized Camelus dromedarius. RNA extracted from 6 healthy female camels was pooled together and used for construction of library in phagemid vector. The VHH inserts were found in 50–70% of the phagemid vector (pMECS vector) cloned in Escherichia coli and the size of the libraries was estimated to be around 105–107cfu. Sequence analysis of VHHs revealed variations in the length of complementary determining regions and amino acid usage resulting in a variety of different paratopes. Further expression of VHH rescued from the library was optimized in prokaryotic system using pET32 expression system. Functional antibodies generated from this naïve library will be explored for affinity against various antigens in further studies.

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