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Custom Nanobody Production

$8,950.00

Item Cat No.: BCNANO

Antibody: Nanobody

Concentration: 1 mg/ml of purified VHH fragment

Application: In vitro and In vivo

Reactivity: Human, Mouse, Rat

BiCell Scientific Inc’s nanobody development service includes immunization of alpaca (Lama pacos), isolation of peripheral blood mononuclear cells (PBMCs), cloning of VHH domain library and screening with yeast display technique.

Nanobody, also known as single-domain antibody (sdAb), is an antibody fragment consisting of a single monomeric variable antibody domain, known as VHH fragment. Nanobody or sdAb cab be engineered from heavy-chain antibodies found in camelids. Heavy-chain antibodies differ from conventional antibodies in that they lack the light chains of conventional antibodies. Their heavy chains consist in CH2 and CH3 domains but not CH1 domain, which are significantly shorter than conventional antibodies.

Nanobodies, ie VHH fragments from sdAb, have very different binding characteristics from conventional antibodies, largely owing to an extended loop in their complementarity-determining region 3 (CDR3). Such a feature not only improves the hydrophilicity and stability of nanobody, but also allow nanobodies to bind to buried sites inside a protein that are not accessible to conventional antibodies.

BiCell Scientific Inc’s nanobody development service utilizes the natural immune system of alpaca (Lama pacos) to generate de novo nanobody sequences from hyperimmunized B cells. These nanobody sequences are then cloned into a yeast display library to facilitate screening and discovery of high-affinity nanobodies.

Nanobody Workflow Chart

Steps Timeline (weeks) Deliverables/Internal testing milestone QC standard
Phase I: Antigen preparation

(peptide antigen,13-19 aa length, will be designed against target protein and its modified site, synthesized and conjugated to KLH.)

2 weeks / COA

 

 

 

 

Phase II: Animal immunization

(one alpaca will be immunized with antigen; conventional protocol – three injections.)

 

 

8-10 weeks 100 ul antiserum ELISA after 3rd injection
Phase III: Cloning nanobody library

(peripheral blood mononuclear cells will be collected when titer reaches 50,000. cDNA from PBMCs will be amplified with nanobody lead primers to clone the entire nanobody library into yeast display vector pYSD.)

 

 

1-2 weeks PBMCs ELISA
Phase IV: Yeast display screening

(pYSD libraries will be transformed into EBY100 YSD yeast strain. Yeast cells with induced expression of nanobody will be sorted with magnetic beads coated with antigen. Antigen-bound yeast cells will be plated to allow forming single colonies. Nanobody sequences from recovered yeast cells will be cloned into E.coli expression plasmids for manufacturing.)

 

3-4 weeks One E.coli clone (to customer); 1mg of purified nanobody from the clone

(BiCell Scientific will bank all remaining clones; customer can request additional clones by paying a service fee)

Magnetic sorting/ FACS

 

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

  • Full sequence of the selected nanobody
  • One E.coli clone expressing nanobody with the highest yield
  • 1 mg of affinity purified nanobody

 


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

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"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

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