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Human Antibody Library Screening with Yeast Display

Left panel illustrates the yeast display system that features a fusion protein presenting the scFv domain of human antibody on the surface of yeast cell. HA tag and Myc tag allow validation of scFv expression. Right panel shows the antigen binding data of yeast cells. Antigen is labeled with FITC and bound with yeast cells. P4 region in top image is shown at the bottom.

2nd Generation Yeast Display System

Left panel: Diagram shows the design of 2nd generation yeast display system featuring a long stalk and a GPI anchor. Middle panel: Yeast cells display an scFv protein that is bound with FITC-labeled antigen. Right panel: FACS graph shows the number of yeast cells displaying the scFv protein bound to FITC-labeled antigen.

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Establishment & Screening of Human Antibody Library (Patient or Healthy Individual)

$10,950.00

Item Cat No.: BCHULB

Antibody: Monoclonal Antibody

Concentration: 1 mg/ml purified IgG

Application: In Vitro and In Vivo

Reactivity: Human

BiCell Scientific’s human antibody library establishment & screening service utilizes a unique and proprietary yeast display platform to experimentally select human IgG molecules from the cloned library of peripheral blood mononuclear cells (PBMCs).

Human peripheral blood mononuclear cells (PBMCs) contain B cells that actively secrete antibodies in response to a variety of diseases such as infection, inflammation, and cancer. Modern DNA or RNA based tumor vaccine can induce effective antibodies only in a small number of patients, which substantiates the need of isolation of therapeutic antibodies from individual patients.

BiCell Scientific Inc’s human antibody library establishment & screening service utilizes the natural immune system of human patients to harvest the antibody sequences from mature B cells. These antibody sequences are then cloned into a yeast display library to facilitate screening and discovery of therapeutic antibodies.

  • Molecular cloning of Fv domain from human IgG heavy chain and light chain into yeast display library vector pBCYD, expressing the scFv form of fused Fv domains from heavy chain and light chain.
  • Transform pBCYD libraries into BJ5465 yeast strain.
  • Harvest yeast cells with induced expression of scFv. Perform antigen based enrichment with magnetic bead sorting and FACS.
  • Recover sorted yeast cells and plate them to form single colonies.
  • Clone the scFv gene sequences from recovered yeast cells into lentiviral expression vector.
  • Transfection of HEK293T cells with lentiviral expression vectors as well as vectors expressing gag–pol, rev and vsv–g.
  • Harvesting and purification of lentivirus from cell culture medium.
  • Transducing naive Sp2/0-Ag14 hybridoma cells with lentivirus encoding human IgG.
  • Clonal selection and stable cell line establishment in 96-well plate.
  • Validation of human IgG antibody expression in 96-well plate against antigen and selection of the highest expressing clone.
  • Affinity chromatography purification with protein A column.

Customers will receive the following deliverables at the end of each project:

  • Fully sequenced Fv domains of heavy chain and light chain of the selected human antibody
  • One stable Sp2/0-Ag14 hybridoma cell line expressing human monoclonal antibody with the highest yield
  • 1 mg of affinity purified human monoclonal antibody

(Customers select the clone of their interest. BiCell Scientific Inc will bank the remaining clones. Additional clones are offered at $950 each).

For Research Use Only. Not for use in clinical diagnostics or therapeutics.

  • Molecular cloning of Fv domain from human IgG heavy chain and light chain into yeast display library vector pBCYD – 2-3 weeks
  • Yeast cell transformation and expression validation  – 2-3 weeks
  • Yeast display and affinity maturation – 3-4 weeks
  • Cloning and sequencing of affinity selected scFv genes from yeast – 2-3 weeks
  • Assembling human IgG into lentiviral expression system – 2-3 weeks
  • Naive Sp2/0-Ag14 hybridoma cell transduction – 1-2 weeks
  • Clonal selection and stable cell line establishment – 3-4 weeks
  • Human antibody expression validation – 2-3 weeks
  • Cell culture expansion and affinity purification – 2-3 weeks

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Julie Craft Van De Weghe, PhD

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"The assay is a homophilic interaction mediated cell adhesion on purified protein (in this case, immobilized purified Pcdhga9 to Pcdhga9 expressed on cell surface). Compared to control, cell adhesion is reduced in the presence of Pcdhga9 monoclonal antibody supernatants!"

Divyesh Joshi, PhD

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"The rabbit hybridoma supernatants of anti-APOBEC3 project are tested positive by ELISA, and we are very happy about it! We previously tried a company, Abclone. Their Project "A" has immune response that is <10,000 titer in antiserum, which would explain why there is no positive mAb after fusion. Their project "B" didn't have any immune response in rabbit."

Harshita B Gupta, PhD.

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