Product: Phage VHH antibody naïve library
Description: VHH antibody library from non-immunized Dromedaries
Quantity: 2x10e12 phage
Source: Peripheral Blood Mononuclear Cells from non-immunized dromedaries.
Number of PBMC cells: 1x10e10
RNA purification: Total RNA was purified with total RNA extraction kit, followed by silica-spin column method. Total RNA was eluted with RNase-free H2O. Quality of RNA was evaluated by OD260/280 ratio (>1.9) and agarose gel electrophoresis. RNA concentration was estimated using formula of 1.0 OD260 = 70ug/µl.
cDNA synthesis: Products of VH and VHH with expected sizes were amplified from the cDNA. Products of VHH were further purified and modified with notI and Pst I sites for cloning into the CV01 phagemid vector.
Library construction: The ligated DNA was transformed into TG1 cells. The library of phage displayed VHHs is obtained after infecting the TG1cells with M13K07 helper phages. Independent clones were randomly picked and DNA inserts of each clone was sequenced. Over 90% of the clones contain inserts representing VHH sequences.