High-affinity lamprey VLRA and VLRB monoclonal antibodies
Lamprey are members of the ancestral vertebrate taxon (jawless fish), which evolved rearranging antigen receptors convergently with the jawed vertebrates. But instead of Ig superfamily domains, lamprey variable lymphocyte receptors (VLRs) consist of highly diverse leucine-rich repeats. Although VLRs...
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| Format: | Article |
| Language: | en_US |
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United States National Academy of Sciences
2010
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| Online Access: | http://hdl.handle.net/1721.1/52559 https://orcid.org/0000-0003-2398-5896 |
| _version_ | 1826210666898259968 |
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| author | Wittrup, Karl Dane Pancer, Zeev Mariuzza, Roy A. Flajnik, Martin F. Xu, Gang Velikovsky, C. Alejandro Tasumi, Satoshi Gai, S. Annie |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Wittrup, Karl Dane Pancer, Zeev Mariuzza, Roy A. Flajnik, Martin F. Xu, Gang Velikovsky, C. Alejandro Tasumi, Satoshi Gai, S. Annie |
| author_sort | Wittrup, Karl Dane |
| collection | MIT |
| description | Lamprey are members of the ancestral vertebrate taxon (jawless fish), which evolved rearranging antigen receptors convergently with the jawed vertebrates. But instead of Ig superfamily domains, lamprey variable lymphocyte receptors (VLRs) consist of highly diverse leucine-rich repeats. Although VLRs represent the only known adaptive immune system not based on Ig, little is known about their antigen-binding properties. Here we report robust plasma VLRB responses of lamprey immunized with hen egg lysozyme and β-galactosidase (β-gal), demonstrating adaptive immune responses against soluble antigens. To isolate monoclonal VLRs, we constructed large VLR libraries from antigen-stimulated and naïve animals in a novel yeast surface-display vector, with the VLR C-terminally fused to the yeast Flo1p surface anchor. We cloned VLRB binders of lysozyme, β-gal, cholera toxin subunit B, R-phycoerythrin, and B-trisaccharide antigen, with dissociation constants up to the single-digit picomolar range, equivalent to those of high-affinity IgG antibodies. We also isolated from a single lamprey 13 anti-lysozyme VLRA clones with affinities ranging from low nanomolar to mid-picomolar. All of these VLRA clones were closely related in sequence, differing at only 15 variable codon positions along the 244-residue VLR diversity region, which augmented antigen-binding affinity up to 100-fold. Thus, VLRs can provide a protective humoral antipathogen shield. Furthermore, the broad range of nominal antigens that VLRs can specifically bind, and the affinities achieved, indicate a functional parallelism between LRR-based and Ig-based antibodies. VLRs may be useful natural single-chain alternatives to conventional antibodies for biotechnology applications. |
| first_indexed | 2024-09-23T14:53:33Z |
| format | Article |
| id | mit-1721.1/52559 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T14:53:33Z |
| publishDate | 2010 |
| publisher | United States National Academy of Sciences |
| record_format | dspace |
| spelling | mit-1721.1/525592022-10-01T23:12:13Z High-affinity lamprey VLRA and VLRB monoclonal antibodies Wittrup, Karl Dane Pancer, Zeev Mariuzza, Roy A. Flajnik, Martin F. Xu, Gang Velikovsky, C. Alejandro Tasumi, Satoshi Gai, S. Annie Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Chemical Engineering Koch Institute for Integrative Cancer Research at MIT Wittrup, Karl Dane Gai, Annie Wittrup, Karl Dane Lamprey are members of the ancestral vertebrate taxon (jawless fish), which evolved rearranging antigen receptors convergently with the jawed vertebrates. But instead of Ig superfamily domains, lamprey variable lymphocyte receptors (VLRs) consist of highly diverse leucine-rich repeats. Although VLRs represent the only known adaptive immune system not based on Ig, little is known about their antigen-binding properties. Here we report robust plasma VLRB responses of lamprey immunized with hen egg lysozyme and β-galactosidase (β-gal), demonstrating adaptive immune responses against soluble antigens. To isolate monoclonal VLRs, we constructed large VLR libraries from antigen-stimulated and naïve animals in a novel yeast surface-display vector, with the VLR C-terminally fused to the yeast Flo1p surface anchor. We cloned VLRB binders of lysozyme, β-gal, cholera toxin subunit B, R-phycoerythrin, and B-trisaccharide antigen, with dissociation constants up to the single-digit picomolar range, equivalent to those of high-affinity IgG antibodies. We also isolated from a single lamprey 13 anti-lysozyme VLRA clones with affinities ranging from low nanomolar to mid-picomolar. All of these VLRA clones were closely related in sequence, differing at only 15 variable codon positions along the 244-residue VLR diversity region, which augmented antigen-binding affinity up to 100-fold. Thus, VLRs can provide a protective humoral antipathogen shield. Furthermore, the broad range of nominal antigens that VLRs can specifically bind, and the affinities achieved, indicate a functional parallelism between LRR-based and Ig-based antibodies. VLRs may be useful natural single-chain alternatives to conventional antibodies for biotechnology applications. 2010-03-12T21:28:30Z 2010-03-12T21:28:30Z 2009-06 2009-04 Article http://purl.org/eprint/type/JournalArticle 1091-6490 0027-8424 http://hdl.handle.net/1721.1/52559 Tasumi, Satoshi et al. “High-affinity lamprey VLRA and VLRB monoclonal antibodies.” Proceedings of the National Academy of Sciences 106.31 (2009): 12891-12896. © 2009 National Academy of Sciences https://orcid.org/0000-0003-2398-5896 en_US http://dx.doi.org/10.1073/pnas.0904443106 Proceedings of the National Academy of Sciences of the United States of America Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf United States National Academy of Sciences PNAS |
| spellingShingle | Wittrup, Karl Dane Pancer, Zeev Mariuzza, Roy A. Flajnik, Martin F. Xu, Gang Velikovsky, C. Alejandro Tasumi, Satoshi Gai, S. Annie High-affinity lamprey VLRA and VLRB monoclonal antibodies |
| title | High-affinity lamprey VLRA and VLRB monoclonal antibodies |
| title_full | High-affinity lamprey VLRA and VLRB monoclonal antibodies |
| title_fullStr | High-affinity lamprey VLRA and VLRB monoclonal antibodies |
| title_full_unstemmed | High-affinity lamprey VLRA and VLRB monoclonal antibodies |
| title_short | High-affinity lamprey VLRA and VLRB monoclonal antibodies |
| title_sort | high affinity lamprey vlra and vlrb monoclonal antibodies |
| url | http://hdl.handle.net/1721.1/52559 https://orcid.org/0000-0003-2398-5896 |
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