Nanobody Production

Nanobodies: One of the hottest new tools in biomedical research.

We have the Alpacas. We can make Nanobodies® for you.

About Nanobody Production


Single domain antibodies (nanobodies) are increasingly becoming popular as reagents for research, for use as therapeutics, and as reagents for diagnostics. This is due, in large part, to their small size, stability, high affinity, high specificity, ease of manipulation, and ease of production. 

Through resources provided by the National Institute of General Medical Sciences, we have developed a nanobody production pipeline. This enables the University of Kentucky's Protein Core to produce these single-domain antibodies at a very reasonable cost. To generate nanobodies, our facility immunizes alpacas with your protein of interest, amplifies the pool of nanobody cDNAs by PCR, and generates a phagemid library, which is screened by panning. All positive clones are sequenced. The nanobodies produced are purified from E. coli and confirmed to bind to your protein.  In many cases, the core can produce your protein of interest by expressing it in E. coli, insect cells, or yeast.  Although nanobodies are very useful for a variety of applications (i.e. for pulling down complexes, for immunohistochemistry, for ELISA's, etc ) they are often not useful for Western blotting.  This follows since nanobodies target conformational epitopes, which can be lost when running an SDS-PAGE gel.  This is in contrast to conventional antibodies that are more likely to target linear epitopes which are retained on SDS-PAGE.

Production of nanobodies in alpacas and other camelids has a distinct advantage over 
other methods in that a much greater diversity in the nanobody pool is achieved offering 
a better chance of obtaining a nanobody with the desired properties, i.e high affinity, unique epitope, optimal kinetics. An example of such diversity is illustrated below with the sequences of three nanobodies we produced in our alpacas against Maltose Binding Protein compared to a reference sequence.

We are now taking orders for producing nanobodies starting in May 2019. To produce a Nanobody to your protein, we will need ~3 mg of purified protein in PBS or HBS. We will deliver 1 mg of your purified and validated nanobody within ~3-4 months. We will also ship you a sequenced bacterial expression vector that can be used for further nanobody production. Typically we identify additional nanobodies for your protein, and these can be purchased for a nominal cost. In addition to your nanobody, a range of derivatives and conjugates are available. For our services and pricing see our services listed below.

To request a nanobody produced to your protein, please fill out and return the nanobody request form and return via email to Christy Maddox (Christy.Maddox@uky.edu) by April 19th, or contact Lou Hersh at lhersh@uky.edu. We will need your protein for nanobody production by April 26th, 2018.

Nanobody Production Request Form

 

 

EXAMPLES OF NANOBODY USES

SUGGESTED READING

Immunization of Alpacas (Lama Pacos) with Protein Antigens and Production of Antigen-specific Single-Domain Antibodies. K. Martin Chow, Sidney W. Whiteheart, Jeffrey R. Smiley, Savita Sharma, Kathy Boaz, Meggie J. Coleman, Alvina Maynard, Louis B. Hersh, Craig W. Vander Kooi. JOVE. 2019 (in press)

Exploiting Nanobodies' Singular Traits. Ingram JR, Schmidt FI, Ploegh HL. Annu Rev Immunol. 2018 Feb 28.

Reporter-nanobody fusions (RANbodies) as versatile, small, sensitive immunohistochemical reagents. Yamagata M, Sanes JR. Proc Natl Acad Sci U S A. 2018 Feb 27;115(9):2126-2131.

Editorial: Single-Domain Antibodies-Biology, Engineering and Emerging Applications. Henry KA, MacKenzie CR. Front Immunol. 2018 Jan 23;9:41.

Nanobodies as novel agents for cancer therapy. Revets H, De Baetselier P, Muyldermans S. Expert Opin Biol Ther. 2005 Jan;5(1):111-24. Review

Nanobodies and their potential applications. Hassanzadeh-Ghassabeh G, Devoogdt N, De Pauw P, Vincke C, Muyldermans S.  Nanomedicine (Lond). 2013 Jun;8(6):1013-26.

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