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Affinity Maturation or Degeneration for Custom Antibody

$12,950.00

Item Cat No.: BCHUAD

Antibody: Monoclonal Antibody

Concentration: 1 mg/ml purified IgG

Application: In Vitro and In Vivo

Reactivity: Human, Mouse, Rat

BiCell Scientific’s affinity maturation or degeneration service utilizes unique and proprietary yeast 2-hybrid platform and yeast display platform to experimentally alter the binding affinities of monoclonal antibodies to suit a wide range of applications, e.g. antibody repurposing, antibody humanization, antibody safety improvement, bispecific antibody binding adjustment and etc.

Monoclonal antibodies have been approved to treat a variety of diseases. The binding characteristics of these therapeutic antibodies can be altered by maturation (gain of affinity) or degeneration (loss of affinity) techniques to confer different functionalities that will be useful to new areas of research and therapeutics. For example, a bispecific antibody would benefit from such an alteration to reduce its binding affinities to each single antigen, thereby acquiring an increased binding specificity to cells carrying both antigens. A therapeutic antibody would benefit from an increase in its binding affinity, thereby reducing the side effects by administrating at a lower effective dose.

BiCell Scientific’s affinity maturation or degeneration service utilizes unique and proprietary yeast 2-hybrid platform and yeast display platform to screen mutations in the the complementarity-determining regions (CDRs) within the Fv domain of both heavy chain and light chain of the antibody molecule, in order to experimentally alter the binding affinities of custom-made or existing therapeutic monoclonal antibodies to suit a wide range of new applications. BiCell Scientific Inc can guarantee a order of magnitude higher or lower affinity improvement for our maturation or degeneration service respectively.

  • Molecular cloning of CDRs (CDR1, CDR2, and CDR3) from IgG heavy chain and light chain into yeast 2-hybrid library vector pY2H series, expressing the scFv form of fused Fv domains from heavy chain and light chain.
  • Transform pY2H libraries (Y2H system) or pYDP libraries (yeast display system) into EBY100 YSD yeast strain.
  • Perform yeast 2-hybrid screen with different growth stringency levels or yeast display screen with magnetic sorting and FACS sorting.
  • Recover stringency tested or 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 gagpol, rev and vsvg.
  • Harvesting and purification of lentivirus from cell culture medium.
  • Transducing naive Sp2/0-Ag14 hybridoma cells with lentivirus encoding engineered IgG.
  • Clonal selection and stable cell line establishment in 96-well plate.
  • Validation of engineered IgG antibody binding characteristics in 96-well plate against antigen and selection of the desired clones with altered binding affinities.
  • Affinity chromatography purification.

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

  • Fully sequenced Fv domains of heavy chain and light chain of the engineered antibody
  • One stable Sp2/0-Ag14 hybridoma cell line expressing engineered monoclonal antibody with the desired binding affinity
  • 1 mg of affinity purified engineered 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 CDRs (CDR1, CDR2, and CDR3) from IgG heavy chain and light chain of into yeast 2-hybrid vector or yeast display vector – 2-3 weeks
  • Yeast cell transformation and expression validation  – 2-3 weeks
  • Yeast 2-hybrid screening or yeast display screening – 3-4 weeks
  • Cloning and sequencing of affinity selected scFv genes from yeast – 2-3 weeks
  • Assembling engineered IgG in 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
  • Engineered antibody expression validation – 2-3 weeks
  • Cell culture expansion and affinity purification – 2-3 weeks

Reviews

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Be the first to review “Affinity Maturation or Degeneration for Custom Antibody”

"I am really impressed with your approach. We tried multiple times previously to create monoclonal and polyclonal antibodies to claudin-2 and MLCK1. We have had limited success generating polyclonals and no success generating monoclonals. You have generated outstanding monoclonals to both. I look forward to continuing to work with you."

Jerrold R. Turner, M.D., Ph.D.

Brigham and Women’s Hospital | Harvard Medical School

"The polyclonal antibody you generated for KIAA0408 is stunning! KIAA0408 is a novel cilium molecule that has never been studied. So, clearly there will be a lot of demand for it as we have discovered a very interesting finding and the story will be published in a high impact journal. I am strongly inclined to generate monoclonal antibody for this protein too and we should think about patenting it."

Univ.-Prof. Jay Gopalakrishnan PhD

Heinrich-Heine-Universität | Universitätsklinikum Düsseldorf

"Your ARL13B antibody works beautifully!!! We’re so happy to have a cilia-specific antibody made in rat! I can send you high resolution images to be posted on your website."

Julie Craft Van De Weghe, PhD

School of Medicine | University of Washington

"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

School of Medicine | Yale University

"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.

School of Medicine | UT Health San Antonio

"We have tested anti mouse T cell antiserum samples from both rabbits you sent to us.

They worked very well! Thank you!"

Victoria Gorbacheva, PhD.

School of Medicine | Cleveland Clinic

Contact us for questions or custom requests!