Naïve Antibody Libraries Construction

Alpha Lifetech Inc. can offer naïve antibody libraries from non-immunized animals and humans based on our mature phage display technology. Alpha Lifetech Inc.'s antibody discovery R&D team is proud to announce that we can develop and generate a library containing 108 - 1010 independent clones.

Introduction Naïve Antibody Library

Naïve antibody libraries are constructed using B cells of unimmunized or healthy donors, normally focusing on the IgM isotype. Naïve libraries are supposedly able to be used for the isolation of mAbs against any antigen.

The naïve repertoire has its unique advantages in generating antibodies against target antigens. Alpha Lifetech can offer the native antibody libraries (scFv, Fab, VHH Formats and customized) from host animals like humans, Goat, Canine, Chicken, Goose, Pig, Alligator, Bovine, Monkey, Donkey, Horse, Ostrich, Hamster, Rabbit, Camel and Llama. Our aim is to understand and meet the demands of different customers and to assist with any upcoming and emerging problems in research work.

Naïve Antibody Library Construction Service

Naïve libraries have been generated by tapping the natural primary (unselected) immune repertoire (non-immunized donors) via cloning of antibodies that recognize a variety of antigens. The rearranged V genes were amplified by the polymerase chain reaction (PCR) from B-cell mRNAs encoding immunoglobulin M (IgM) taken from non-immunized donors. By using this procedure, antibodies were recovered before an encounter with antigen and unscreened for tolerance by the immune system. Indeed, a naïve library represents a good source of Abs against self, non-immunogenic, and toxic antigens if the library is sufficiently large and diverse.

To create a large-size antibody library, Alpha Lifetech Inc. utilizes an efficient two-step cloning strategy for the construction of antibody phage libraries based on the isolation of restriction fragments from plasmid vectors instead of PCR products. When digesting PCR products with restriction sites defined by the oligonucleotide primer, efficiency is dependent on the number of extra nucleotides appended to the primer, but it is always low compared to the digestion of plasmid DNA. In the first step of the procedure described, primary repertoires are prepared from PCR products encoding the VH, Vκ, and Vλ domains, yielding typical medium-sized libraries (1 - 100 million clones). In the second step, VH fragments are isolated by digestion of plasmid DNA purified from the primary repertoires and cloned into the acceptor phagemid vector containing the light-chain (LC) repertoires. This innovation increases the size of libraries dramatically (10 - 100 billion clones).

The general developmental procedures of naïve antibody phage display libraries are shown in the figure: we collect the peripheral blood lymphocytes (PBLs) from non-immunized hosts (human, different animals), separate the antibody gene of interest and then clone into phagemid gene fragment of interest and form a phage display library with a specific antibody fragment.

Why Choose Us?

-- Excellent for the Production of Naïve Antibodies in Any Species

-- Construction of Naïve Fab, scFv, and VHH Libraries

-- Isolation of Human Monoclonal Antibody

-- Selecting Antibodies to Cell-Surface/Tumor-associated Antigens 
 
If you have any questions, please feel free to contact us at any time.