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Rational development of high-affinity T-cell receptor-like antibodies.

ABSTRACT: T-cell interaction with a target cell is a key event in the adaptive immune response and primarily driven by T-cell receptor (TCR) recognition of peptide-MHC (pMHC) complexes. TCR avidity for a given pMHC is determined by number of MHC molecules, availability of coreceptors, and TCR affinity for MHC or peptide, respectively, with peptide recognition being the most important factor to confer target specificity. Here we present high-resolution crystal structures of 2 Fab antibodies in complex with the immunodominant NY-ESO-1(157-165) peptide analogue (SLLMWITQV) presented by HLA-A*0201 and compare them with a TCR recognizing the same pMHC. Binding to the central methionine-tryptophan peptide motif and orientation of binding were almost identical for Fabs and TCR. As the MW "peg" dominates the contacts between Fab and peptide, we estimated the contributions of individual amino acids between the Fab and peptide to provide the rational basis for a peptide-focused second-generation, high-affinity antibody library. The final Fab candidate achieved better peptide binding by 2 light-chain mutations, giving a 20-fold affinity improvement to 2-4 nM, exceeding the affinity of the TCR by 1,000-fold. The high-affinity Fab when grafted as recombinant TCR on T cells conferred specific killing of HLA-A*0201/NY-ESO-1(157-165) target cells. In summary, we prove that affinity maturation of antibodies mimicking a TCR is possible and provide a strategy for engineering high-affinity antibodies that can be used in targeting specific pMHC complexes for diagnostic and therapeutic purposes.

SUBMITTER: Stewart-Jones G

PROVIDER: S-EPMC2667008 | biostudies-literature | 2009 Apr

REPOSITORIES: biostudies-literature

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Rational development of high-affinity T-cell receptor-like antibodies.

Stewart-Jones Guillaume G Wadle Andreas A Hombach Anja A Shenderov Eugene E Held Gerhard G Fischer Eliane E Kleber Sascha S Nuber Natko N Stenner-Liewen Frank F Bauer Stefan S McMichael Andrew A Knuth Alexander A Abken Hinrich H Hombach Andreas A AA Cerundolo Vincenzo V Jones E Yvonne EY Renner Christoph C

Proceedings of the National Academy of Sciences of the United States of America 20090323 14

T-cell interaction with a target cell is a key event in the adaptive immune response and primarily driven by T-cell receptor (TCR) recognition of peptide-MHC (pMHC) complexes. TCR avidity for a given pMHC is determined by number of MHC molecules, availability of coreceptors, and TCR affinity for MHC or peptide, respectively, with peptide recognition being the most important factor to confer target specificity. Here we present high-resolution crystal structures of 2 Fab antibodies in complex with ...[more]

PMID: 19307587

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Project description:Targeting the CD47-signal-regulatory protein ? (SIRP?) pathway represents a novel therapeutic approach to enhance anti-cancer immunity by promoting both innate and adaptive immune responses. Unlike CD47, which is expressed ubiquitously, SIRP? expression is mainly restricted to myeloid cells and neurons. Therefore, compared to CD47-targeted therapies, targeting SIRP? may result in differential safety and efficacy profiles, potentially enabling lower effective doses and improved pharmacokinetics and pharmacodynamics. The development of effective SIRP? antagonists is restricted by polymorphisms within the CD47-binding domain of SIRP?, necessitating pan-allele reactive anti-SIRP? antibodies for therapeutic intervention in diverse patient populations. We immunized wild-type and human antibody transgenic chickens with a multi-allele and multi-species SIRP? regimen in order to discover pan-allelic and pan-mammalian reactive anti-SIRP? antibodies suitable for clinical translation. A total of 200 antibodies were isolated and screened for SIRP? reactivity from which approximately 70 antibodies with diverse SIRP? binding profiles, sequence families, and epitopes were selected for further characterization. A subset of anti-SIRP? antibodies bound to both human SIRP? v1 and v2 alleles with high affinity ranging from low nanomolar to picomolar, potently antagonized the CD47/SIRP? interaction, and potentiated macrophage-mediated antibody-dependent cellular phagocytosis in vitro. X-ray crystal structures of five anti-SIRP? antigen-binding fragments, each with unique epitopes, in complex with SIRP? (PDB codes 6NMV, 6NMU, 6NMT, 6NMS, and 6NMR) are reported. Furthermore, some of the anti-SIRP? antibodies cross-react with cynomolgus SIRP? and various mouse SIRP? alleles (BALB/c, NOD, BL/6), which can facilitate preclinical to clinical development. These properties provide an attractive rationale to advance the development of these anti-SIRP? antibodies as a novel therapy for advanced malignancies. Abbreviations: ADCC: antibody-dependent cellular cytotoxicity; ADCP: antibody-dependent cellular phagocytosis; CFSE: carboxyfluorescein succinimidyl ester; Fab: fragment antigen binding; Fc: fragment crystallizable; Fc?R: Fc? receptor; Ig: immunoglobulin; IND: investigational new drug; MDM?: monocyte-derived macrophage; NOD: non-obese diabetic; scFv: single chain fragment variable; SCID: severe combined immunodeficiency; SIRP: signal-regulatory protein.

Dataset Information

Rational development of high-affinity T-cell receptor-like antibodies.

Publications

Rational development of high-affinity T-cell receptor-like antibodies.

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