Nanobody Services

New in January 2022!

The CMI has launched Yeast Surface Display Nanobody Selection Services, based on libraries developed in the lab of Andrew Kruse. 

To start a conversation about a nanobody project, contact the CMI.

 

CMI Nanobody Selection Terms of Service (for Academic Users)

The CMI charges for labor and supplies to perform nanobody selection campaigns, but not for commercial licensing.  Academic labs are free to use nanobodies, clones or sequences identified in a CMI selection campaign for academic purposes.

External Academic Labs

A Nanobody Services Agreement and a Purchase Order are required of all external academic labs prior project initiation. 

CMI Academic User Agreement for Nanobody Services, sample user agreement for for External Academic Labs requesting Yeast Surface Display Nanobody Selections.  A new agreement is required for each selection project. Do not complete or sign the Sample form. The CMI will provide you with a clean form after consulting with you regarding the scope of your project.

CMI nanobody selections are performed using yeast surface display nanobody libraries developed in the Kruse Laboratory and is the property of Harvard University.  The terms outlined in the CMI Academic User Agreement for Nanobody Services apply to any external academic labs receiving the Kruse lab Nanobody Library or clones selected from the library, from any source, and outlines a revenue sharing agreement in the event that compounds derived from the library are commercialized.  

Note to Industry Users: we will not be offering Nanobody selection services to commercial institutions at the launch.  You may request more information on commercial licensing opportunities by contacting the CMI or the Kruse Lab.

CMI Nanobody Services

Currently, the CMI is offering a basic yeast display selection service and  a set of additional nanobody services.

Basic Yeast Display Nanobody Service

nanobody basic service

Additional Nanobody Services

Nanobody Additional Services

Nanobody Selection Sample Requirements

Method Notes Buffer Notes minimum volume (μl)

recommended minimum
concentration (mg/ml)

Nanobody Selection Campaign (4 rounds) per antigen, standard selection antigen buffer needs to be free of primary amines for chemical biotinylation 1 ml 15 μM

About Nanobodies

The majority of antibodies used in the laboratory are of mammalian origin and have a conventional bivalent architecture consisting of two heavy chains and two light chains. The heavy and light chains have terminal variable domains responsible for antigen specificity, VHand VL. While camelids also produce conventional antibodies, the majority of circulating antibodies have a different architecture, consisting only of two heavy chains, called heavy chain antibodies.The smaller variable domains of camelid antibodies (VHH domains), also called nanobodies, allow them to bind in clefts of folded proteins that are not accessible by larger heterodimeric Fab and single-chain variable domain (scFv) fragments. Nanobodies have been particularly useful as tools to facilitate structural studies and to modulate function of many proteins including integral membrane receptors.Unlike conventional antibodies or antibody fragments which have multiple obligate interchain and intrachain disulfide bonds, the camelid VHH domain framework (which can have up to two disulfides), generally retains function in the reducing environment of the cytoplasm. Combined with their small size, this makes nanobodies particularly useful tools for intracellular applications such as super-resolution live-cell imaging.3,4 In addition to being useful laboratory tools, nanobodies are plausible scaffolds for drug development. Nanobody scaffolds have been humanizedand the first nanobody drug (Caplacizumab) was approved by the FDA.6

antibody structure

While camelid heavy chain antibodies can be produced by immunization of camels, llamas or alpacas, the process requires a large animal husbandry facility and is time consuming and expensive. The Kruse lab has developed a synthetic yeast display library based on a consensus framework derived from llama antibody genes with variable complementary determining region loops (CDRs) designed from known nanobody structures.The laboratory is developing additional, second generation libraries. We plan to initiate the CMI antibody discovery platform by offering nanobody production services based on these unique HMS resources. 

Antibodies derived using surface display technologies, such as yeast display, offer several unique features rarely accessible by animal immunization.The most practical advantage of surface display technologies is that antibody fragments are sequence-verified and renewable, allowing for rapid subcloning into a variety of expression systems for production in many formats. Specificity can be expanded or restricted by including secondary or counter screens with additional antigens. In vitro selection methods have been used to achieve selective recognition of chemical modifications and proteins with single surface-exposed amino acid differences, where conventional immunization has failed. In vitro selection methods avoid immunological tolerance, which can restrict the production of antibodies produced in mammals against highly conserved antigenic targets. Affinity maturation to improve affinity or selectivity, by generation of secondary libraries of mutagenized variants, can be performed at any stage using the same selection process. Surface display methods can also be used to directly select human antibody frameworks, avoiding the need to humanize animal frameworks for conversion to biologics. 

 

  1. Hamers-Casterman, C. et al.Naturally occurring antibodies devoid of light chains. Nature363,446–448 (1993).
  2. Manglik, A., Kobilka, B. K. & Steyaert, J. Nanobodies to Study G Protein-Coupled Receptor Structure and Function. Annu. Rev. Pharmacol. Toxicol.57,19–37 (2017).
  3. Pleiner, T. et al.Nanobodies: site-specific labeling for super-resolution imaging, rapid epitope-mapping and native protein complex isolation. Elife4,e11349 (2015).
  4. Schumacher, D., Helma, J., Schneider, A. F. L., Leonhardt, H. & Hackenberger, C. P. R. Nanobodies: Chemical Functionalization Strategies and Intracellular Applications. Angew. Chem. Int. Ed. Engl.57,2314–2333 (2018).
  5. Vincke, C. et al.General strategy to humanize a camelid single-domain antibody and identification of a universal humanized nanobody scaffold. J Biol Chem284,3273–3284 (2009).
  6. Kaplon, H. & Reichert, J. M. Antibodies to watch in 2019. MAbs11,219–238 (2019).
  7. McMahon, C. et al.Yeast surface display platform for rapid discovery of conformationally selective nanobodies. Nat Struct Mol Biol25,289–296 (2018).
  8. Bradbury, A. R. M., Sidhu, S., Dübel, S. & McCafferty, J. Beyond natural antibodies: the power of in vitro display technologies. Nat Biotechnol29,245–254 (2011).
  9. Wörn, A. & Pluckthun, A. Stability engineering of antibody single-chain Fv fragments. J Mol Biol305,989–1010 (2001).

FY23 Nanobody Selection Fees

Please contact the CMI to start a conversation about nanobody selections.  Fees for Nanobody selections cover supplies and labor and are assessed for completed stages in the selection, regardless of final experimental outcome of the selection. 

Service

Harvard Internal

(33-digit)

External
Academic
Antigen Purification by SEC
Required if not completed by user
$650 $793.00
Yeast Surface Display Selection Campaign
Includes 2 rounds MACS, 2 rounds FACS and clonal isolation
delivery of any sequences or clones produced
$7980 $9735.60
Optional - Expression and Purification of Nanobody
1L scale, in e.coli pET26b, with 6-His tag
$4180 $5099.60
Total
includes selection campaign and nanobody expression
$12810 $15628.20