1lmk › Citations

Crystal structure of a diabody, a bivalent antibody fragment.

Perisic O, Webb PA, Holliger P, Winter G, Williams RL
Structure 2 1217-26 (1994)
Cited: 116 times
EuropePMC logo PMID: 7704531

Abstract

Background

Diabodies are dimeric antibody fragments. In each polypeptide, a heavy-chain variable domain (VH) is linked to a light-chain variable domain (VL) but unlike single-chain Fv fragments, each antigen-binding site is formed by pairing of one VH and one VL domain from the two different polypeptides. Diabodies thus have two antigen-binding sites, and can be bispecific. Direct structural evidence is lacking for the connections and dimeric interactions between the two polypeptides of the diabody.

Results

The 2.6 A resolution structure has been determined for a bivalent diabody with a flexible five-residue polypeptide linker between the (amino-terminal) VH and (carboxy-terminal) VL domains. The asymmetric unit of the crystal consists of four polypeptides comprising two diabodies; for one of these polypeptides the linker can be traced between the VH and VL domains. Within each diabody the two associated VH and VL domains make back-to-back interactions through the VH domains, and there is an extensive VL-VL interface between the two diabodies in the asymmetric unit.

Conclusion

The structure of the diabody is very similar to that which had been predicted by molecular modelling. Diabodies directed against cell-surface antigens should be capable of bringing together two cells, such as in cell-targeted therapy, because the two antigen-binding sites of the diabody are at opposite ends of the molecule and separated by approximately 65 A.

Articles - 1lmk mentioned but not cited (12)

  1. Examination of the contributions of size and avidity to the neutralization mechanisms of the anti-HIV antibodies b12 and 4E10. Klein JS, Gnanapragasam PN, Galimidi RP, Foglesong CP, West AP, Bjorkman PJ. Proc Natl Acad Sci U S A 106 7385-7390 (2009)
  2. Antigen-independent activation enhances the efficacy of 4-1BB-costimulated CD22 CAR T cells. Singh N, Frey NV, Engels B, Barrett DM, Shestova O, Ravikumar P, Cummins KD, Lee YG, Pajarillo R, Chun I, Shyu A, Highfill SL, Price A, Zhao L, Peng L, Granda B, Ramones M, Lu XM, Christian DA, Perazzelli J, Lacey SF, Roy NH, Burkhardt JK, Colomb F, Damra M, Abdel-Mohsen M, Liu T, Liu D, Standley DM, Young RM, Brogdon JL, Grupp SA, June CH, Maude SL, Gill S, Ruella M. Nat Med 27 842-850 (2021)
  3. Structure-function analysis of a lupus anti-DNA autoantibody: central role of the heavy chain complementarity-determining region 3 Arg in binding of double- and single-stranded DNA. Li Z, Schettino EW, Padlan EA, Ikematsu H, Casali P. Eur J Immunol 30 2015-2026 (2000)
  4. Development of PF-06671008, a Highly Potent Anti-P-cadherin/Anti-CD3 Bispecific DART Molecule with Extended Half-Life for the Treatment of Cancer. Root AR, Root AR, Cao W, Li B, LaPan P, Meade C, Sanford J, Jin M, O'Sullivan C, Cummins E, Lambert M, Sheehan AD, Ma W, Gatto S, Kerns K, Lam K, D'Antona AM, Zhu L, Brady WA, Benard S, King A, He T, Racie L, Arai M, Barrett D, Stochaj W, LaVallie ER, Apgar JR, Svenson K, Mosyak L, Yang Y, Chichili GR, Liu L, Li H, Burke S, Johnson S, Alderson R, Finlay WJJ, Lin L, Olland S, Somers W, Bonvini E, Gerber HP, May C, Moore PA, Tchistiakova L, Bloom L. Antibodies (Basel) 5 E6 (2016)
  5. Association of putative concave protein-binding sites with the fluctuation behavior of residues. Ertekin A, Nussinov R, Haliloglu T. Protein Sci 15 2265-2277 (2006)
  6. Crystal structures of mono- and bi-specific diabodies and reduction of their structural flexibility by introduction of disulfide bridges at the Fv interface. Kim JH, Song DH, Youn SJ, Kim JW, Cho G, Kim SC, Lee H, Jin MS, Lee JO. Sci Rep 6 34515 (2016)
  7. Molecular basis for the antagonistic activity of an anti-CXCR4 antibody. Peng L, Damschroder MM, Cook KE, Wu H, Dall'Acqua WF. MAbs 8 163-175 (2016)
  8. Diabody-Ig: a novel platform for the generation of multivalent and multispecific antibody molecules. Seifert O, Rau A, Beha N, Richter F, Kontermann RE. MAbs 11 919-929 (2019)
  9. The enigma of the near-symmetry of proteins: Domain swapping. Bonjack-Shterengartz M, Avnir D. PLoS One 12 e0180030 (2017)
  10. The sclerostin-neutralizing antibody AbD09097 recognizes an epitope adjacent to sclerostin's binding site for the Wnt co-receptor LRP6. Boschert V, Frisch C, Back JW, van Pee K, Weidauer SE, Muth EM, Schmieder P, Beerbaum M, Knappik A, Timmerman P, Mueller TD. Open Biol 6 160120 (2016)
  11. Distinguishing Between Monomeric scFv and Diabody in Solution Using Light and Small Angle X-ray Scattering. Lüdel F, Bufe S, Bleymüller WM, de Jonge H, Iamele L, Niemann HH, Hellweg T. Antibodies (Basel) 8 E48 (2019)
  12. Structural arrangement of the VH and VL domains in the COBRA™ T-cell engaging single-chain diabody. Krakow J, Hammel M, Zhu Y, Hillier BJ, Paolella B, Desmarais A, Wall R, Chen TT, Pei R, Karunatilake C, DuBridge R, Vinogradova M. Antib Ther 5 1-10 (2022)


Reviews citing this publication (18)

  1. 3D domain swapping: as domains continue to swap. Liu Y, Eisenberg D. Protein Sci 11 1285-1299 (2002)
  2. The making of bispecific antibodies. Brinkmann U, Kontermann RE. MAbs 9 182-212 (2017)
  3. Antibody constructs in cancer therapy: protein engineering strategies to improve exposure in solid tumors. Beckman RA, Weiner LM, Davis HM. Cancer 109 170-179 (2007)
  4. Strategies for selection of antibodies by phage display. Griffiths AD, Duncan AR. Curr Opin Biotechnol 9 102-108 (1998)
  5. CAR T cells for brain tumors: Lessons learned and road ahead. Akhavan D, Alizadeh D, Wang D, Weist MR, Shepphird JK, Brown CE. Immunol Rev 290 60-84 (2019)
  6. Antibody engineering. Maynard J, Georgiou G. Annu Rev Biomed Eng 2 339-376 (2000)
  7. Dimeric and trimeric antibodies: high avidity scFvs for cancer targeting. Kortt AA, Dolezal O, Power BE, Hudson PJ. Biomol Eng 18 95-108 (2001)
  8. High avidity scFv multimers; diabodies and triabodies. Hudson PJ, Kortt AA. J Immunol Methods 231 177-189 (1999)
  9. Immunoglobulin domains in Escherichia coli and other enterobacteria: from pathogenesis to applications in antibody technologies. Bodelón G, Palomino C, Fernández LÁ. FEMS Microbiol Rev 37 204-250 (2013)
  10. Expanding the Boundaries of Biotherapeutics with Bispecific Antibodies. Husain B, Ellerman D. BioDrugs 32 441-464 (2018)
  11. Bispecific antibodies: molecules that enable novel therapeutic strategies. Fischer N, Léger O. Pathobiology 74 3-14 (2007)
  12. Generation and production of engineered antibodies. Kipriyanov SM, Le Gall F. Mol Biotechnol 26 39-60 (2004)
  13. Single-Chain Fragment Variable: Recent Progress in Cancer Diagnosis and Therapy. Muñoz-López P, Ribas-Aparicio RM, Becerra-Báez EI, Fraga-Pérez K, Flores-Martínez LF, Mateos-Chávez AA, Luria-Pérez R. Cancers (Basel) 14 4206 (2022)
  14. Minimalist Antibodies and Mimetics: An Update and Recent Applications. Bruce VJ, Ta AN, McNaughton BR. Chembiochem 17 1892-1899 (2016)
  15. Production and structure of diabodies. Poljak RJ. Structure 2 1121-1123 (1994)
  16. Clinical genetics for the pulmonologist: introduction. Cottin V. Respiration 74 3-7 (2007)
  17. Monoclonal antibodies and antibody fragments: state of the art and future perspectives in the treatment of non-haematological tumors. Di Fede G, Bronte G, Rizzo S, Rolfo Cervetto C, Cocorullo G, Gulotta G, Bazan V, Russo A. Expert Opin Biol Ther 11 1433-1445 (2011)
  18. Structural Analysis of Recent Allergen-Antibody Complexes and Future Directions. Mueller GA, Min J, Foo ACY, Pomés A, Pedersen LC. Curr Allergy Asthma Rep 19 17 (2019)

Articles citing this publication (86)

  1. Beyond natural antibodies: the power of in vitro display technologies. Bradbury AR, Sidhu S, Dübel S, McCafferty J. Nat Biotechnol 29 245-254 (2011)
  2. Conformations of the third hypervariable region in the VH domain of immunoglobulins. Morea V, Tramontano A, Rustici M, Chothia C, Lesk AM. J Mol Biol 275 269-294 (1998)
  3. Optimizing the stability of single-chain proteins by linker length and composition mutagenesis. Robinson CR, Sauer RT. Proc Natl Acad Sci U S A 95 5929-5934 (1998)
  4. Tumor localization of anti-CEA single-chain Fvs: improved targeting by non-covalent dimers. Wu AM, Chen W, Raubitschek A, Williams LE, Neumaier M, Fischer R, Hu SZ, Odom-Maryon T, Wong JY, Shively JE. Immunotechnology 2 21-36 (1996)
  5. Bispecific tandem diabody for tumor therapy with improved antigen binding and pharmacokinetics. Kipriyanov SM, Moldenhauer G, Schuhmacher J, Cochlovius B, Von der Lieth CW, Matys ER, Little M. J Mol Biol 293 41-56 (1999)
  6. Prolonged in vivo tumour retention of a human diabody targeting the extracellular domain of human HER2/neu. Adams GP, Schier R, McCall AM, Crawford RS, Wolf EJ, Weiner LM, Marks JD. Br J Cancer 77 1405-1412 (1998)
  7. Tuning cytokine receptor signaling by re-orienting dimer geometry with surrogate ligands. Moraga I, Wernig G, Wilmes S, Gryshkova V, Richter CP, Hong WJ, Sinha R, Guo F, Fabionar H, Wehrman TS, Krutzik P, Demharter S, Plo I, Weissman IL, Minary P, Majeti R, Constantinescu SN, Piehler J, Garcia KC. Cell 160 1196-1208 (2015)
  8. A revival of bispecific antibodies. Kufer P, Lutterbüse R, Baeuerle PA. Trends Biotechnol 22 238-244 (2004)
  9. Single-chain antibody displayed on a recombinant measles virus confers entry through the tumor-associated carcinoembryonic antigen. Hammond AL, Plemper RK, Zhang J, Schneider U, Russell SJ, Cattaneo R. J Virol 75 2087-2096 (2001)
  10. Crystal structures of NK1-heparin complexes reveal the basis for NK1 activity and enable engineering of potent agonists of the MET receptor. Lietha D, Chirgadze DY, Mulloy B, Blundell TL, Gherardi E. EMBO J 20 5543-5555 (2001)
  11. Di-, tri- and tetrameric single chain Fv antibody fragments against human CD19: effect of valency on cell binding. Le Gall F, Kipriyanov SM, Moldenhauer G, Little M. FEBS Lett 453 164-168 (1999)
  12. High level secretion of a humanized bispecific diabody from Escherichia coli. Zhu Z, Zapata G, Shalaby R, Snedecor B, Chen H, Carter P. Biotechnology (N Y) 14 192-196 (1996)
  13. Triabodies: single chain Fv fragments without a linker form trivalent trimers. Iliades P, Kortt AA, Hudson PJ. FEBS Lett 409 437-441 (1997)
  14. The 2.0-A resolution crystal structure of a trimeric antibody fragment with noncognate VH-VL domain pairs shows a rearrangement of VH CDR3. Pei XY, Holliger P, Murzin AG, Williams RL. Proc Natl Acad Sci U S A 94 9637-9642 (1997)
  15. scFv multimers of the anti-neuraminidase antibody NC10: length of the linker between VH and VL domains dictates precisely the transition between diabodies and triabodies. Atwell JL, Breheney KA, Lawrence LJ, McCoy AJ, Kortt AA, Hudson PJ. Protein Eng 12 597-604 (1999)
  16. Bispecific CD3 x CD19 diabody for T cell-mediated lysis of malignant human B cells. Kipriyanov SM, Moldenhauer G, Strauss G, Little M. Int J Cancer 77 763-772 (1998)
  17. A bispecific diabody directed against prostate-specific membrane antigen and CD3 induces T-cell mediated lysis of prostate cancer cells. Bühler P, Wolf P, Gierschner D, Schaber I, Katzenwadel A, Schultze-Seemann W, Wetterauer U, Tacke M, Swamy M, Schamel WW, Elsässer-Beile U. Cancer Immunol Immunother 57 43-52 (2008)
  18. The structure of a polyQ-anti-polyQ complex reveals binding according to a linear lattice model. Li P, Huey-Tubman KE, Gao T, Li X, West AP, Bennett MJ, Bjorkman PJ. Nat Struct Mol Biol 14 381-387 (2007)
  19. The crystal structure of an anti-CEA scFv diabody assembled from T84.66 scFvs in V(L)-to-V(H) orientation: implications for diabody flexibility. Carmichael JA, Power BE, Garrett TP, Yazaki PJ, Shively JE, Raubischek AA, Wu AM, Hudson PJ. J Mol Biol 326 341-351 (2003)
  20. Inhibition of papillomavirus protein function in cervical cancer cells by intrabody targeting. Griffin H, Elston R, Jackson D, Ansell K, Coleman M, Winter G, Doorbar J. J Mol Biol 355 360-378 (2006)
  21. Fab-based bispecific antibody formats with robust biophysical properties and biological activity. Wu X, Sereno AJ, Huang F, Lewis SM, Lieu RL, Weldon C, Torres C, Fine C, Batt MA, Fitchett JR, Glasebrook AL, Kuhlman B, Demarest SJ. MAbs 7 470-482 (2015)
  22. Redirected optimized cell killing (ROCK®): A highly versatile multispecific fit-for-purpose antibody platform for engaging innate immunity. Ellwanger K, Reusch U, Fucek I, Wingert S, Ross T, Müller T, Schniegler-Mattox U, Haneke T, Rajkovic E, Koch J, Treder M, Tesar M. MAbs 11 899-918 (2019)
  23. Effect of domain order on the activity of bacterially produced bispecific single-chain Fv antibodies. Kipriyanov SM, Moldenhauer G, Braunagel M, Reusch U, Cochlovius B, Le Gall F, Kouprianova OA, Von der Lieth CW, Little M. J Mol Biol 330 99-111 (2003)
  24. The effect of variable domain orientation and arrangement on the antigen-binding activity of a recombinant human bispecific diabody. Lu D, Jimenez X, Witte L, Zhu Z. Biochem Biophys Res Commun 318 507-513 (2004)
  25. Phage diabody repertoires for selection of large numbers of bispecific antibody fragments. McGuinness BT, Walter G, FitzGerald K, Schuler P, Mahoney W, Duncan AR, Hoogenboom HR. Nat Biotechnol 14 1149-1154 (1996)
  26. The importance of framework residues H6, H7 and H10 in antibody heavy chains: experimental evidence for a new structural subclassification of antibody V(H) domains. Jung S, Spinelli S, Schimmele B, Honegger A, Pugliese L, Cambillau C, Plückthun A. J Mol Biol 309 701-716 (2001)
  27. On the origin of the histone fold. Alva V, Ammelburg M, Söding J, Lupas AN. BMC Struct Biol 7 17 (2007)
  28. IL-8 single-chain homodimers and heterodimers: interactions with chemokine receptors CXCR1, CXCR2, and DARC. Leong SR, Lowman HB, Liu J, Shire S, Deforge LE, Gillece-Castro BL, McDowell R, Hébert CA. Protein Sci 6 609-617 (1997)
  29. Optimized linker sequences for the expression of monomeric and dimeric bispecific single-chain diabodies. Völkel T, Korn T, Bach M, Müller R, Kontermann RE. Protein Eng 14 815-823 (2001)
  30. Bispecific anti-CD20/22 antibodies inhibit B-cell lymphoma proliferation by a unique mechanism of action. Qu Z, Goldenberg DM, Cardillo TM, Shi V, Hansen HJ, Chang CH. Blood 111 2211-2219 (2008)
  31. Construction and characterization of a bispecific diabody for retargeting T cells to human carcinomas. Helfrich W, Kroesen BJ, Roovers RC, Westers L, Molema G, Hoogenboom HR, de Leij L. Int J Cancer 76 232-239 (1998)
  32. Complement recruitment using bispecific diabodies. Kontermann RE, Wing MG, Winter G. Nat Biotechnol 15 629-631 (1997)
  33. Crystal structure of the anti-(carcinoembryonic antigen) single-chain Fv antibody MFE-23 and a model for antigen binding based on intermolecular contacts. Boehm MK, Corper AL, Wan T, Sohi MK, Sutton BJ, Thornton JD, Keep PA, Chester KA, Begent RH, Perkins SJ. Biochem J 346 Pt 2 519-528 (2000)
  34. Expression of a bispecific dsFv-dsFv' antibody fragment in Escherichia coli. Schmiedl A, Breitling F, Dübel S. Protein Eng 13 725-734 (2000)
  35. Engineered humanized diabodies for microPET imaging of prostate stem cell antigen-expressing tumors. Leyton JV, Olafsen T, Sherman MA, Bauer KB, Aghajanian P, Reiter RE, Wu AM. Protein Eng Des Sel 22 209-216 (2009)
  36. Design and Characterization of a Human Monoclonal Antibody that Modulates Mutant Connexin 26 Hemichannels Implicated in Deafness and Skin Disorders. Xu L, Carrer A, Zonta F, Qu Z, Ma P, Li S, Ceriani F, Buratto D, Crispino G, Zorzi V, Ziraldo G, Bruno F, Nardin C, Peres C, Mazzarda F, Salvatore AM, Raspa M, Scavizzi F, Chu Y, Xie S, Yang X, Liao J, Liu X, Wang W, Wang S, Yang G, Lerner RA, Mammano F. Front Mol Neurosci 10 298 (2017)
  37. Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments. Lawrence LJ, Kortt AA, Iliades P, Tulloch PA, Hudson PJ. FEBS Lett 425 479-484 (1998)
  38. Enzyme immunoassays using bispecific diabodies. Kontermann RE, Martineau P, Cummings CE, Karpas A, Allen D, Derbyshire E, Winter G. Immunotechnology 3 137-144 (1997)
  39. One size does not fit all: navigating the multi-dimensional space to optimize T-cell engaging protein therapeutics. Chen W, Yang F, Wang C, Narula J, Pascua E, Ni I, Ding S, Deng X, Chu ML, Pham A, Jiang X, Lindquist KC, Doonan PJ, Van Blarcom T, Yeung YA, Chaparro-Riggers J. MAbs 13 1871171 (2021)
  40. Reversible dimer formation and stability of the anti-tumour single-chain Fv antibody MFE-23 by neutron scattering, analytical ultracentrifugation, and NMR and FT-IR spectroscopy. Lee YC, Boehm MK, Chester KA, Begent RH, Perkins SJ. J Mol Biol 320 107-127 (2002)
  41. Three-dimensional structures of single-chain Fv-neuraminidase complexes. Malby RL, McCoy AJ, Kortt AA, Hudson PJ, Colman PM. J Mol Biol 279 901-910 (1998)
  42. A highly effective and stable bispecific diabody for cancer immunotherapy: cure of xenografted tumors by bispecific diabody and T-LAK cells. Hayashi H, Asano R, Tsumoto K, Katayose Y, Suzuki M, Unno M, Kodama H, Takemura S, Yoshida H, Makabe K, Imai K, Matsuno S, Kumagai I, Kudo T. Cancer Immunol Immunother 53 497-509 (2004)
  43. A potent antagonist antibody targeting connexin hemichannels alleviates Clouston syndrome symptoms in mutant mice. Kuang Y, Zorzi V, Buratto D, Ziraldo G, Mazzarda F, Peres C, Nardin C, Salvatore AM, Chiani F, Scavizzi F, Raspa M, Qiang M, Chu Y, Shi X, Li Y, Liu L, Shi Y, Zonta F, Yang G, Lerner RA, Mammano F. EBioMedicine 57 102825 (2020)
  44. Construction and humanization of a functional bispecific EGFR × CD16 diabody using a refolding system. Asano R, Nakayama M, Kawaguchi H, Kubota T, Nakanishi T, Umetsu M, Hayashi H, Katayose Y, Unno M, Kudo T, Kumagai I. FEBS J 279 223-233 (2012)
  45. Therapeutic potential of an anti-HER2 single chain antibody-DM1 conjugates for the treatment of HER2-positive cancer. Zhang H, Wang Y, Wu Y, Jiang X, Tao Y, Yao Y, Peng Y, Chen X, Fu Y, Yu L, Wang R, Lai Q, Lai W, Li W, Kang Y, Yi S, Lu Y, Gou L, Wu M, Yang J. Signal Transduct Target Ther 2 17015 (2017)
  46. VH/VL interface engineering to promote selective expression and inhibit conformational isomerization of thrombopoietin receptor agonist single-chain diabody. Igawa T, Tsunoda H, Kikuchi Y, Yoshida M, Tanaka M, Koga A, Sekimori Y, Orita T, Aso Y, Hattori K, Tsuchiya M. Protein Eng Des Sel 23 667-677 (2010)
  47. Exploiting the avian immunoglobulin system to simplify the generation of recombinant antibodies to allergenic proteins. Finlay WJ, deVore NC, Dobrovolskaia EN, Gam A, Goodyear CS, Slater JE. Clin Exp Allergy 35 1040-1048 (2005)
  48. A Structurally Unresolved Head Segment of Defined Length Favors Proper Measles Virus Hemagglutinin Tetramerization and Efficient Membrane Fusion Triggering. Navaratnarajah CK, Rosemarie Q, Cattaneo R. J Virol 90 68-75 (2016)
  49. Acquired antagonistic activity of a bispecific diabody directed against two different epitopes on vascular endothelial growth factor receptor 2. Lu D, Kotanides H, Jimenez X, Zhou Q, Persaud K, Bohlen P, Witte L, Zhu Z. J Immunol Methods 230 159-171 (1999)
  50. Highly enhanced cytotoxicity of a dimeric bispecific diabody, the hEx3 tetrabody. Asano R, Ikoma K, Sone Y, Kawaguchi H, Taki S, Hayashi H, Nakanishi T, Umetsu M, Katayose Y, Unno M, Kudo T, Kumagai I. J Biol Chem 285 20844-20849 (2010)
  51. Intracellular and cell surface displayed single-chain diabodies. Kontermann RE, Müller R. J Immunol Methods 226 179-188 (1999)
  52. Single-chain Fv multimers of the anti-neuraminidase antibody NC10: the residue at position 15 in the V(L) domain of the scFv-0 (V(L)-V(H)) molecule is primarily responsible for formation of a tetramer-trimer equilibrium. Dolezal O, De Gori R, Walter M, Doughty L, Hattarki M, Hudson PJ, Kortt AA. Protein Eng 16 47-56 (2003)
  53. Crystal structures of a rat anti-CD52 (CAMPATH-1) therapeutic antibody Fab fragment and its humanized counterpart. Cheetham GM, Hale G, Waldmann H, Bloomer AC. J Mol Biol 284 85-99 (1998)
  54. The role of valency in the selection of anti-carbohydrate single-chain Fvs from phage display libraries. MacKenzie R, To R. J Immunol Methods 220 39-49 (1998)
  55. A single mutation in framework 2 of the heavy variable domain improves the properties of a diabody and a related single-chain antibody. Rodríguez-Rodríguez ER, Ledezma-Candanoza LM, Contreras-Ferrat LG, Olamendi-Portugal T, Possani LD, Becerril B, Riaño-Umbarila L. J Mol Biol 423 337-350 (2012)
  56. Construction of a diabody (small recombinant bispecific antibody) using a refolding system. Takemura S, Asano R, Tsumoto K, Ebara S, Sakurai N, Katayose Y, Kodama H, Yoshida H, Suzuki M, Imai K, Matsuno S, Kudo T, Kumagai I. Protein Eng 13 583-588 (2000)
  57. Factory neovessels: engineered human blood vessels secreting therapeutic proteins as a new drug delivery system. Compte M, Alonso-Camino V, Santos-Valle P, Cuesta AM, Sánchez-Martín D, López MR, Vicario JL, Salas C, Sanz L, Alvarez-Vallina L. Gene Ther 17 745-751 (2010)
  58. Application of the Fc fusion format to generate tag-free bi-specific diabodies. Asano R, Ikoma K, Kawaguchi H, Ishiyama Y, Nakanishi T, Umetsu M, Hayashi H, Katayose Y, Unno M, Kudo T, Kumagai I. FEBS J 277 477-487 (2010)
  59. A trivalent anti-erbB2/anti-CD16 bispecific antibody retargeting NK cells against human breast cancer cells. Xie Z, Shi M, Feng J, Yu M, Sun Y, Shen B, Guo N. Biochem Biophys Res Commun 311 307-312 (2003)
  60. Crystal structures of a quorum-quenching antibody. Debler EW, Kaufmann GF, Kirchdoerfer RN, Mee JM, Janda KD, Wilson IA. J Mol Biol 368 1392-1402 (2007)
  61. Rapid phenotyping of HPA-1a using either diabody-based hemagglutination or recombinant IgG1-based assays. Watkins NA, Armour KL, Smethurst PA, Metcalfe P, Scott ML, Hughes DL, Smith GA, Williamson LM, Clark MR, Ouwehand WH. Transfusion 39 781-789 (1999)
  62. A diabody that dissociates to monomer forms at low concentration: effects on binding activity and tumor targeting. Huang BC, Foote LJ, Lankford TK, Davern SM, McKeown CK, Kennel SJ. Biochem Biophys Res Commun 327 999-1005 (2005)
  63. A Human-Derived Monoclonal Antibody Targeting Extracellular Connexin Domain Selectively Modulates Hemichannel Function. Ziraldo G, Buratto D, Kuang Y, Xu L, Carrer A, Nardin C, Chiani F, Salvatore AM, Paludetti G, Lerner RA, Yang G, Zonta F, Mammano F. Front Physiol 10 392 (2019)
  64. Structural considerations for functional anti-EGFR × anti-CD3 bispecific diabodies in light of domain order and binding affinity. Asano R, Nagai K, Makabe K, Takahashi K, Kumagai T, Kawaguchi H, Ogata H, Arai K, Umetsu M, Kumagai I. Oncotarget 9 13884-13893 (2018)
  65. Bispecific antibodies for treatment of cancer in experimental animal models and man. de Leij L, Molema G, Helfrich W, Kroesen BJ. Adv Drug Deliv Rev 31 105-129 (1998)
  66. Bispecific targeting of thrombin activatable fibrinolysis inhibitor and plasminogen activator inhibitor-1 by a heterodimer diabody. Develter J, Booth NA, Declerck PJ, Gils A. J Thromb Haemost 6 1884-1891 (2008)
  67. Characterisation of a panel of anti-tetanus toxin single-chain Fvs reveals cooperative binding. Scott N, Qazi O, Wright MJ, Fairweather NF, Deonarain MP. Mol Immunol 47 1931-1941 (2010)
  68. Mono and bivalent binding of a scFv and covalent diabody to murine laminin-1 using radioiodinated proteins and SPR measurements: effects on tissue retention in vivo. Huang BC, Davern S, Kennel SJ. J Immunol Methods 313 149-160 (2006)
  69. A semi high-throughput method for screening small bispecific antibodies with high cytotoxicity. Sugiyama A, Umetsu M, Nakazawa H, Niide T, Onodera T, Hosokawa K, Hattori S, Asano R, Kumagai I. Sci Rep 7 2862 (2017)
  70. Bivalent Fv antibody fragments obtained by substituting the constant domains of a fab fragment with heterotetrameric molybdopterin synthase. Petrov K, Dion M, Hoffmann L, Dintinger T, Defontaine A, Tellier C. J Mol Biol 341 1039-1048 (2004)
  71. Preferential heterodimerization of a bispecific diabody based on a humanized anti-EGFR antibody 528. Asano R, Sone Y, Ikoma K, Hayashi H, Nakanishi T, Umetsu M, Katayose Y, Unno M, Kudo T, Kumagai I. Protein Eng Des Sel 21 597-603 (2008)
  72. Effective screening method of agonistic diabodies based on autocrine growth. Nakano K, Kojima T, Kasutani K, Senoh C, Natori O, Ishii S, Tsunoda H, Hattori K. J Immunol Methods 347 31-35 (2009)
  73. Small molecule AX-024 reduces T cell proliferation independently of CD3ϵ/Nck1 interaction, which is governed by a domain swap in the Nck1-SH3.1 domain. Richter K, Rufer AC, Muller M, Burger D, Casagrande F, Grossenbacher T, Huber S, Hug MN, Koldewey P, D'Osualdo A, Schlatter D, Stoll T, Rudolph MG. J Biol Chem 295 7849-7864 (2020)
  74. An antibody loop replacement design feasibility study and a loop-swapped dimer structure. Clark LA, Boriack-Sjodin PA, Day E, Eldredge J, Fitch C, Jarpe M, Miller S, Li Y, Simon K, van Vlijmen HW. Protein Eng Des Sel 22 93-101 (2009)
  75. Evaluation of three different formats of a neutralizing single chain human antibody against toxin Cn2: neutralization capacity versus thermodynamic stability. Quintero-Hernández V, Del Pozo-Yauner L, Pedraza-Escalona M, Juárez-González VR, Alcántara-Recillas I, Possani LD, Becerril B. Immunol Lett 143 152-160 (2012)
  76. Regulation of the flavin redox potential by flavin-binding antibodies. Bruggeman YE, Honegger A, Kreuwel H, Visser AJ, Laane C, Schots A, Hilhorst R. Eur J Biochem 249 393-400 (1997)
  77. Crystal structure of an anti-interleukin-2 monoclonal antibody Fab complexed with an antigenic nonapeptide. Afonin PV, Fokin AV, Tsygannik IN, Mikhailova IY, Onoprienko LV, Mikhaleva II, Ivanov VT, Mareeva TY, Nesmeyanov VA, Li N, Pangborn WA, Duax WL, Pletnev VZ. Protein Sci 10 1514-1521 (2001)
  78. Domain swap in the C-terminal ubiquitin-like domain of human doublecortin. Rufer AC, Kusznir E, Burger D, Stihle M, Ruf A, Rudolph MG. Acta Crystallogr D Struct Biol 74 450-462 (2018)
  79. Antibody gene transfer treatment drastically improves epidermal pathology in a keratitis ichthyosis deafness syndrome model using male mice. Peres C, Sellitto C, Nardin C, Putti S, Orsini T, Di Pietro C, Marazziti D, Vitiello A, Calistri A, Rigamonti M, Scavizzi F, Raspa M, Zonta F, Yang G, White TW, Mammano F. EBioMedicine 89 104453 (2023)
  80. Different tissue distribution properties for glycosylation variants of fusion proteins containing the p40 subunit of murine interleukin-12. Bootz F, Venetz D, Ziffels B, Neri D. Protein Eng Des Sel 29 445-455 (2016)
  81. Structure of an influenza neuraminidase-diabody complex by electron cryomicroscopy and image analysis. Rosenthal PB, Waddington LJ, Hudson PJ. J Mol Biol 334 721-731 (2003)
  82. Chemically Crosslinked Bispecific Antibodies for Cancer Therapy: Breaking from the Structural Restrictions of the Genetic Fusion Approach. Ueda A, Umetsu M, Nakanishi T, Hashikami K, Nakazawa H, Hattori S, Asano R, Kumagai I. Int J Mol Sci 21 E711 (2020)
  83. Construction of non-covalent single-chain Fv dimers for hepatocellular carcinoma and their biological functions. Bie CQ, Yang DH, Liang XJ, Tang SH. World J Hepatol 2 185-191 (2010)
  84. Cross-linking disulfide bonds govern solution structures of diabodies. Math BA, Waibl F, Lamp LM, Fernández-Quintero ML, Liedl KR. Proteins 91 1316-1328 (2023)
  85. Modulating Antibody Structure and Function through Directed Mutations and Chemical Rescue. Kaiser CE, Rincon Pabon JP, Khowsathit J, Castaldi MP, Kazmirski SL, Weis DD, Zhang AX, Karanicolas J. ACS Synth Biol 7 1152-1162 (2018)
  86. Production and characterization of a bacterial single-chain Fv fragment specific to human truncated midkine. Dansithong W, Paul S, Mitsumoto T, Saruhashi S, Shinozawa T. Cancer Lett 164 169-176 (2001)


Quick links