To start a conversation about a nanobody project, contact the CMI.
Please contact the CMI to start a conversation about nanobody selections.
CMI nanobody selections are performed using yeast surface display nanobody libraries developed in the Laboratory of Andrew Kruse, which are the property of Harvard University.
Academic labs are free to use nanobodies, clones or sequences identified in a CMI selection campaign without restrictions for academic purposes.
Fees are assessed for completed stages in the selection, regardless of final experimental outcome of the selection. Users will be consulted about progress and advised on whether the selection should proceed after each phase of the selection.
Note to Industry Users: we do not offer Nanobody selection services to commercial institutions at this time. You may request more information on commercial licensing opportunities by contacting the CMI or the Kruse Lab directly.
Currently, the CMI is offering basic yeast display selection services and a set of additional optional nanobody services. Successful campaigns typically yeild 5-20 unique clones with expressable clones having affinities in the 100 nM to uM range, from the native library.
For Nanobody selections, the CMI currently uses a synthetic yeast display library, developed in the Kruse lab, a based on a consensus framework derived from llama antibody genes with variable complementary determining region loops (CDRs) designed from known structures of nanobodies against protein antigens.1
McMahon, C. et al.Yeast surface display platform for rapid discovery of conformationally selective nanobodies. Nat Struct Mol Biol25,289–296 (2018).
Method | Notes | Buffer Notes | minimum volume |
recommended minimum |
---|---|---|---|---|
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 |
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.1 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.2 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 humanized5 and the first nanobody drug (Caplacizumab) was approved by the FDA.6
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.7 The CMI antibody discovery platform currently offers nanobody production services based on this unique HMS resource.
Antibodies derived using surface display technologies, such as yeast display, offer several unique features rarely accessible by animal immunization.8 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.
To learn more about nanobody selections at the CMI, visit our Nanobody Services page.
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.
Harvard base price | External Academic (base+F&A) | |
Antigen Purification by SEC | $660 | $792 |
Required, if not performed by user | ||
Basic Yeast Surface Display Nanobody Selection Campaign | $8,650 | $10,380 |
Includes 2 rounds MACS, 2 rounds FACS, clonal isolation, NGS of enriched library, delivery of any sequences or clones produced. | ||
Full Yeast Surface Display Nanobody Selection Campaign | $9,840 | $11,808 |
Includes Basic Yeast Surface Display Selection Campaign plus cloning into pET26b for periplasmic bacterial expression, for up to 20 isolated clones. | ||
Nanobody Expression and Purification | $4,410 | $5,292 |
Requires Full Yeast Surface Display Selection. Includes small scale periplasmic expression, purification and FSEC, then large scale express and purification of up to 3 Nb, if soluble. |
To start a conversation about a nanobody project, contact the CMI.