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Modification of a human monoclonal antibody Fab fragment specific for Plasmodium falciparum 19-kDa C-terminal merozoite surface protein 1 by site-directed mutagenesis

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Abstract

We recently produced human monoclonal antibody Fab fragments specific for the 19-kDa C-terminal merozoite surface protein 1 of Plasmodium falciparum in a bacterial expression system. The effect of single amino acid modifications in the third complementarity-determining regions of the heavy and light chains on affinity was examined in one of the Fab fragments, Pf25. Recombination polymerase chain reaction was used to modify Tyr92 or Ile97 in the light chain and Val101 or Trp107 in the heavy chain. No effective replacements for Tyr92 and Val101 were found, but possible substitutions of Ile97 with Gly, Leu, Glu, Ala and Ser, and of Trp107 with Arg and Ser were demonstrated. Of these modified Fab fragments, the affinities of Fabs with Ile97–Leu and Trp107–Ser mutations were slightly higher than that of the original Fab. The effects of these modifications on the antigen–antibody interaction are discussed.

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Acknowledgments

This work was supported by a grant-in-aid for scientific research from the Japanese Society for the Promotion of Science and grants from the Ministry of Health, Labor, and Welfare of Japan (to H. T.).

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Authors and Affiliations

  1. Department of Medical Microbiology and Parasitology, Fudan University School of Medicine, Shanghai, 200032, China

    Yan-Lin Tao, Xun-Jia Cheng & Yong-Feng Fu

  2. Teaching and Research Support Center, Tokai University School of Medicine, Isehara, Kanagawa, 259-1193, Japan

    Hideo Tsukamoto

  3. Department of Molecular Life Sciences, Tokai University School of Medicine, Isehara, Kanagawa, 259-1193, Japan

    Eisaku Yoshihara

  4. Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa, 259-1193, Japan

    Xun-Jia Cheng & Hiroshi Tachibana

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  1. Yan-Lin Tao
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  2. Xun-Jia Cheng
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  3. Yong-Feng Fu
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  4. Hideo Tsukamoto
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  5. Eisaku Yoshihara
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  6. Hiroshi Tachibana
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Correspondence to Hiroshi Tachibana.

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Tao, YL., Cheng, XJ., Fu, YF. et al. Modification of a human monoclonal antibody Fab fragment specific for Plasmodium falciparum 19-kDa C-terminal merozoite surface protein 1 by site-directed mutagenesis. Parasitol Res 103, 429–433 (2008). https://doi.org/10.1007/s00436-008-0994-4

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  • DOI: https://doi.org/10.1007/s00436-008-0994-4

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