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. 1994 Mar;85(3):298–305. doi: 10.1111/j.1349-7006.1994.tb02097.x

Development and Characterization of Chimeric Anti‐carcinoembryonic Antigen Monoclonal Antibodies and Their Fab Fragments

Takashi Kamigaki 1,✉, Harumasa Ohyanagi 2, Masahiro Yamamoto 1, Teruo Kaneda 3, Takashi Goto 4, Takao Ohmura 4, Kazumasa Yokoyama 4, Yoichi Saitoh 1
PMCID: PMC5919441  PMID: 8188529

Abstract

In an attempt to reduce the immunogenicity of two different murine anti‐carcinoembryonic antigen (CEA) monoclonal antibodies (MAbs), KM10 and A10, we produced recombinant mouse/human chimeric MAbs and the respective Fab fragments carrying the variable regions of the murine MAbs. Chimeric A10 Fab fragment was expressed in Escherichia coli, and produced in large quantities in a mini‐jar fermentation system. In competitive binding assays, chimeric MAbs and their Fab fragments showed identical specificity to human CEA epitopes, as compared to the parental MAbs or Fab fragments. Both chimeric Fab fragments exhibited strong immunohistochemical reactivity with various gastrointestinal carcinomas and no reactivity with CEA‐related antigens, such as NCA (nonspecific cross‐reacting antigen) or BGPI (biliary glycoprotein I). Furthermore, chimeric KM10 MAb elicited substantially higher antibody‐dependent cellular cytotoxicity than the murine MAb. Complement‐dependent cytotoxicity in vitro was much weaker with chimeric KM10 MAb. These results indicate that chimeric MAbs or Fab fragments could potentially replace the parental murine antibodies or their Fab fragments in therapy or diagnosis of human gastrointestinal carcinomas.

Keywords: Chimeric monoclonal antibody, Chimeric Fab fragment, KM10, A10, Carcinoembryonic antigen

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REFERENCES

  • 1. ) Mach , J. P. , Buchegger , F. , Forni , M. , Ritschard , J. , Berche , C. , Lumbroso , J. D. , Schreyer , M. , Girardet , C. , Accola , R. S. and Carrel , S.Use of radiolabeled monoclonal anti‐CEA antibodies for the detection of human carcinomas by external photoscanning and tomoscintigraphy . Immunol. Today , 2 , 239 – 249 ( 1981. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Dillman , R. O. , Shawler , D. L. , Sobol , R. E. , Collins , H. A. , Beauregard , J. C. , Wormsley , S. B. and Royston , I.Murine monoclonal antibody therapy in two patients with chronic lymphocytic leukemia . Blood , 59 , 1036 – 1045 ( 1982. ). [PubMed] [Google Scholar]
  • 3. ) Larson , S. M. , Carrasquillo , J. A. , Krohn , K. A. , Brown , J. P. , Guffin , R. W. , Ferens , J. M. , Graham , M. M. , Hill , L. D. , Beaumier , P. L. and Hellstrom , K. E.Localization of 131I‐labeled p97‐specific Fab fragments in human melanoma as a basis for radiotherapy . J. Clin. Invest. , 72 , 2101 – 2114 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. ) Ohyanagi , H. , Ishida , H. , Ishida , T. , Soyama , N. , Yamamoto , M. , Okumura , S. , Kano , Y. , Ueda , Y. and Saitoh , Y.A monoclonal antibody KM10 reactive with human gastrointestinal cancer and its application for immunotherapy . Jpn. J. Cancer Res. , 79 , 1349 – 1358 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. ) Ishida , H. , Ohyanagi , H. , Okumura , S. and Saitoh , Y.A monoclonal antibody KM10 reactive with human gastrointestinal cancer . J. Jpn. Surg. Soc. , 89 , 508 – 515 ( 1988. ) ( in Japanese ). [PubMed] [Google Scholar]
  • 6. ) Soyama , N. , Yamamoto , M. , Ohyanagi , H. and Saitoh , Y.Comparative studies on monoclonal antibody KM10 and anti‐CEA monoclonal antibodies . J. Jpn. Surg. Soc. , 90 , 1834 – 1839 ( 1989. ) ( in Japanese ). [PubMed] [Google Scholar]
  • 7. ) Schroff , R. W. , Foon , K. A. , Beatty , S. M. , Oldham , R. K. and Morgan , A. C. , Jr.Human anti‐murine immunoglobulin response in patients receiving monoclonal antibody therapy . Cancer Res. , 45 , 879 – 885 ( 1985. ). [PubMed] [Google Scholar]
  • 8. ) Shawler , D. L. , Bartholomew , R. M. , Smith , L. M. and Dillman , R. O.Human immune response to multiple injections of murine monoclonal IgG1 . J. Immunol. , 135 , 1530 – 1535 ( 1985. ). [PubMed] [Google Scholar]
  • 9. ) Courtency‐Luck , N. S. , Epenetos , A. A. , Moore , R. , Larche , M. , Pectasides , D. , Dhokia , B. and Ritter , M. A.Development of primary and secondary immune response to mouse monoclonal antibodies used in the diagnosis and therapy of malignant neoplasms . Cancer Res. , 46 , 6489 – 6493 ( 1986. ). [PubMed] [Google Scholar]
  • 10. ) Better , M. , Chang , C. P. , Robinson , R. R. and Horwitz , A. H.Escherichia coli secretion of an active chimeric antibody fragment . Science , 240 , 1041 – 1043 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 11. ) Johnston , S. , Lee , J. H. and Ray , D. S.High‐level expression of M13 gene II protein from an inducible polycistronic messenger RNA . Gene , 34 , 137 – 145 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Uemura , Y. , Fukuyama , K. , Nishida , M. , Suyama , Y. and Ohori , H.Establishment of passive hemagglutination assay (PHA) system for anti‐HBc in plasma . Tohoku J. Exp. Med. , 149 , 11 – 20 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Soyama , N. , Ohyanagi , H. and Saitoh , Y.Production of human monoclonal antibody reactive with gastrointestinal carcinoma . Jpn. J. Gastroenterol. , 87 , 2635 – 2641 ( 1990. ) ( in Japanese ). [PubMed] [Google Scholar]
  • 14. ) Morrison , S. , Johnson , M. J. , Herzenberg , L. A. and Oi , V. T.Chimeric human antibody molecules: mouse antigen‐binding domains with human constant region domains . Proc. Natl. Acad. Sci. USA , 81 , 6851 – 6855 ( 1984. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. ) Boulianne , G. L. , Hozumi , N. and Shulman , M. J.Production of functional chimeric mouse/human antibody . Nature , 312 , 643 – 646 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Buchegger , F. , Pelegrin , A. , Hardman , N. , Heusser , C. , Lukas , J. , Dolci , W. and Mach , J. P.Different behaviour of mouse‐human chimeric antibody F(ab')2 fragments of IgG1, IgG2 and IgG4 sub‐class in vivo . Int. J. Cancer , 50 , 416 – 422 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) Siegall , C. B. , Gawlak , S. L. , Chin , J. J. , Zoeckler , M. E. , Kadow , K. F. , Brown , J. P. and Braslawsky , G. R.Cytotoxicity of chimeric (human‐mouse) monoclonal antibody BR96 IgG, F(ab')2 and Fab' conjugated to Pseudomonas exotoxin . Bioconjugate Chem. , 3 , 302 – 307 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Skerra , A. and Plückthun , A.Assembly of a functional immunoglobulin Fv fragment in Escherichia coli . Science , 240 , 1038 – 1041 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 19. ) Horwitz , A. H. , Chang , C. P. , Better , M. , Hellstrom , K. E. and Robinson , R. R.Secretion of functional antibody and Fab fragment from yeast cells . Proc. Natl. Acad. Sci. USA , 85 , 8678 – 8682 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. ) von Kleist , S. , Chavanel , G. and Burtin , P.Identification of an antigen from normal human tissue that crossreacts with the carcinoembryonic antigen . Proc. Natl. Acad. Sci. USA , 69 , 2492 – 2494 ( 1972. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. ) Bordes , M. , Knobel , S. and Martin , F.Carcinoembryonic antigen and related antigens in blood cells and hematopoietic tissues . Eur. J. Cancer , 11 , 783 – 786 ( 1975. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Svenberg , T.Carcinoembryonic antigen‐like substances of human bile. Isolation and partial characterization . Int. J. Cancer , 17 , 588 – 596 ( 1976. ). [DOI] [PubMed] [Google Scholar]
  • 23. ) Svenberg , T. , Hammarström , S. and Hedin , A.Purification and properties of biliary glycoprotein (BGPI). Immunochemical relationship to carcinoembryonic antigen . Mol. Immunol. , 16 , 245 – 252 ( 1979. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) Nap , M. , Hammarström , M. L. , Bormer , O. , Hammarström , S. , Wagener , C. , Handt , S. , Schreyer , M. , Mach , J. P. , Buchegger , F. , von Kleist , S. , Grunert , F. , Seguin , P. , Fuks , A. , Holm , R. and Lamerz , R.Specificity and affinity of monoclonal antibodies against carcinoembryonic antigen . Cancer Res. , 15 , 2329 – 2339 ( 1992. ). [PubMed] [Google Scholar]
  • 25. ) Brüggemann , M. , Williams , G. T. , Bindon , C. I. , Clark , M. R. , Walker , M. R. , Jeffris , R. , Waldmann , H. and Neuberger , M. S.Comparison of the effector function of human immunoglobulins using a matched set of chimeric antibodies . J. Exp. Med. , 166 , 1351 – 1361 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26. ) Steplewski , Z. , Sun , L. E. , Shearman , C. W. , Ghrayeb , J. , Daddona , P. and Koprowski , H.Biological activity of human‐mouse IgG1, IgG2, IgG3, and IgG4 chimeric monoclonal antibodies with antitumor specificity . Proc. Natl. Acad. Sci. USA , 85 , 4852 – 4856 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27. ) Wilbanks , T. , Peterson , J. A. , Miller , S. , Kaufman , L. , Ortendahl , D. and Ceriani , R. L.Localization of mammary tumors in vivo with 131I‐labeled Fab fragment of antibodies against mouse mammary epithelial (MME) antigens . Cancer , 48 , 1768 – 1775 ( 1981. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Beaumier , P. L. , Krohn , K. A. , Carrasquillo , J. A. , Eary , J. , Hellström , I. , Hellström , K. E. , Nelp , W. B. and Larson , S. M.Melanoma localization in nude mice with monoclonal Fab against p97 . J. Nucl. Med. , 26 , 1172 – 1179 ( 1985. ). [PubMed] [Google Scholar]
  • 29. ) LoBuglio , A. F. , Wheeler , R. H. , Trang , J. , Haynes , A. , Rogers , K. , Harvey , E. B. , Sun , L. , Ghrayeb , J. and Khazaeli , M. B.Mouse/human chimeric monoclonal antibody in man: kinetics and immune response . Proc. Natl. Acad. Sci. USA , 86 , 4220 – 4224 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]

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