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De Novo Nanobody Discovery & Maturation

$7,450.00

Item Cat No.: BCNALI

Antibody: Nanobody

Concentration: 1 mg/ml of purified VHH fragment

Application: In vitro and In vivo

Reactivity: Human, Mouse, Rat

BiCell Scientific’s de novo nanobody discovery & maturation service utilizes a unique and proprietary yeast display platform to experimentally select nanobody molecules from a synthetic alpaca (Lama pacos) VHH library.

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’s synthetic nanobody library is constructed from a consensus framework derived from 105 unique nanobody sequences in the PDB database. The constant framework is combined with designed variations in the CDRs that comprise the highly variable antigen-binding interface. Randomization of residue composition in the CDRs is achieved by using a mixed pool of timer phosphoramidites to match target codon frequency. The resulting synthetic antibody library contains >1 × 108 unique antibody clones that are able to be expressed and displayed on the yeast cell surface. By using MACS and FACS enrichment techniques, BiCell Scientific’s de novo nanobody discovery & maturation service can guarantee the selection of nanobodies with binding affinity of EC50 value at 100 nM or less.

  • Construct yeast display library vector pYSD series, expressing the single monomeric variable antibody domain, VHH fragment.
  • Transform pYSD libraries into EBY100 YSD yeast strain.
  • Harvest yeast cells with induced expression of VHH. Perform antigen based binding enrichment with MACS and FACS.
  • Recover sorted yeast cells and plate them to form single colonies.
  • Clone the VHH gene sequences from recovered yeast cells into E.coli expression vector.
  • Transform E.coli cells and induce nanobody expression with IPTG.
  • Affinity purify nanobody with Nickel column and gel filtration column.

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

(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.

  • Initial screening of yeast display library against antigen protein – 3-4 weeks
  • One round of MACS enrichment and maturation  – 2-3 weeks
  • One round of FACS enrichment and maturation – 2-3 weeks
  • Cloning and sequencing of affinity selected VHH genes from yeast – 2-3 weeks
  • Cloning VHH genes into E.coli expression vector – 1-2 weeks
  • E.coli transformation and expression of nanobody – 2-3 weeks
  • Affinity purification and gel filtration – 2-3 weeks

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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

"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

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