Yeast dispaly based Naïve VHH Library from Camel

Product Detail

High-diversity large naïve VHH library from Camel for antibody discovery. Yeast display from Camel enables screening of recombinant single-domain antibodies (VHHs) with subnanomolar affinity, suitable for FACS, ELISA, WB, and therapeutic research applications.

Yeast display or yeast surface display is a novel technique widely used to express the proteins at the yeast surface after translation and maturation in a eukaryotic system. Thus, the yeast display is superior to phage display for display of protein or antibody because the eukaryotic expression environment of yeast cells ensures fine tuning such as folding, modification, and translocation prior to display on the surface.

The preparation of single domain antibody libraries starting from lymphocytes recovered from immunized members of the Camelidae enables to collect binders that underwent somatic maturation. However, the time and costs necessary to prepare a library for each new antigen may urge to look for alternatives such as those offered by large one-pot libraries. Here we constructed four large naïve libraries using PBMCs and spleens from 20 healthy nonimmunized Bactrian camels.The library consists of more than 10⁹ individual transformants with more than 90 % unique sequences and the positive insert ratio was more than 99.99%. We leverage this established library along with our advanced screening platform to extend the epitope coverage and help clients find the optimal monoclonal antibody candidates Despite the lack of somatic maturation, the selection based on yeast display allowed to isolate from such naïve libraries VHHs with affinity in the subnanomolar range and suitable for the standard immunoapplications.

Library Overview

Library Type: Yeast display based Naive VHH Library from Camel
Species Source: Camel
Diversity: >10⁹ individual transformants with >90% unique sequences
Positive Insert Rate: >99.99%
Format: 1 mL ready-to-use library

Yeast dispaly based Naïve VHH Library from Camel

Key Features of Yeast Display

Direct connection between genotype and phenotype
Each yeast cell displays 10⁴–10⁵ copies of VHH
Allows quantitative measurement of binding and expression via FACS
Enables 3–5 rounds of enrichment for high-affinity clones
Compatible with recombinant camelid VHH library screening

Similar to phage display, yeast display provides a direct connection between genotype and phenotype. A plasmid containing the gene of interest is contained within yeast cells and the encoded VHH is expressed on the cell surface.

The display level of each yeast cell is variable, with each cell displaying 10⁴ to 10⁵ copies of the VHHs.

Screening of VHH variation of surface expression and avidity can be quantified using fluorescence-activated cell sorting (FACS), which measures both the strength of antigen binding and amount of VHH expression on the yeast cell surface using separate tags on the antibody and antigen.

FACS binding assays provide a much more quantitative way of assessing high binding affinity and selectivity for the antigens and offer the ability to accurately control selection parameters (binding population percentage, signal normalization, and desired binding affinities) by flow cytometry.

There are usually 3-5 rounds of enrichment of target antigen-binding clones.

The VHHs displayed by a single yeast colony is evaluated for specificity and reproducibility. If the VHH has the required functionality, the antibody gene is sequenced as part of antibody validation.

Screening & Affinity Selection

Fluorescence-activated cell sorting (FACS) enables selection based on binding affinity and expression level
Affinity in the subnanomolar range achieved from naïve libraries
Provides reproducible and validated VHH candidates for immunoassays and therapeutic research
Naive VHH Library from Camel ensures broad epitope coverage for diverse antigen targets

Applications

Application	Description
Antibody Screening	Identify monoclonal VHH candidates using yeast display from Camel
Flow Cytometry	Quantitative assessment of VHH binding via FACS
Immunoassays	ELISA, Western blot (WB), Immunoprecipitation (IP) validation
Therapeutic Research	Discovery of high-affinity single-domain antibodies for drug development