Efficient one-step direct labelling of recombinant antibodies with technetium-99m (original) (raw)

Abstract

High-affinity bacterially expressed antibody fragments can nowadays be cloned from established hybridomas or, more conveniently, isolated directly from antibody libraries displayed on filamentous phage. Such antibodies can be tagged with C-terminal peptide tags containing one cysteine residue, which represents a convenient functionalisation site for a number of applications, including technetium-99m labelling. Here we describe a simple one-step method for99m-Tc labelling of cysteine-tagged recombinant antibodies with more than 50% radionuclide incorporation. The labelled antibodies displayed full retention of immuoreactivity and good stability.

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References

  1. Winter G, Griffiths AD, Hawkins RE, Hoogenboom HR. Making antibodies by phage display technology.Annu Rev Immunol 1994; 12:433–455.
    Google Scholar
  2. Yokota T, Milenic DE, Whitlow M, Wood JF, Hubert SL, Schlom J. Microautoradiographic analysis of the normal organ distributions of radioiodinated scFv and other immunoglobulin forms.Cancer Res 1993; 53: 3776–3783.
    Google Scholar
  3. Verharr MJ, Keep PA, Hewkins RE, Robson L, Redley B, Boden JA, Begent RHJ, Chester KA.Int J Cancer 1995; 61: 497–501.
    Google Scholar
  4. Griffiths AD, Williams SC, Hartley O, Tomlinson IM, Waterhouse P, Crosby WL, Kontermann RE, Jones PT, Low NM, Allison TJ, Prospero TD, Hoogenboom HR, Nissim A, Cox JPL, Harrison JL, Zaccolo M, Gherardi E, Winter G. Isolation of high affinity human antibodies directly from large synthetic repertoires.EMBO J 1994; 13: 3245–3260.
    Google Scholar
  5. Neri D, Petrul H, Roncucci G. Engineering recombinant antibodies for immunotherapy.Cell Biophys 1995; 27: 47–61.
    Google Scholar
  6. Cumber AJ, Ward ES, Winter G, Parnell GD, Warzynczak EJ. Comparative stabilities in vitro and in vivo of a recombinant mouse antibody FvCys fragment and a bisFvCys conjugate.J Immunol 1992; 149: 120–126.
    Google Scholar
  7. Neri D, Momo M, Prospero T, Winter G. High-affinity antigen binding by chelating recombinant antibodies (CRAbs).J Mol Biol 1995; 246: 367–373.
    Google Scholar
  8. Hoogenboom HR, Griffiths AD, Johnson KS, Chiswell DJ, Hudson P, Winter G. Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains.Nucleic Acids Res 1991; 19: 4133–4137.
    Google Scholar
  9. King DJ, Turner A, Farnsworth AP, Adair JR, Owens RJ, Pedley RB, Baldock D, Proudfoot KA, Lawson ADG, Beeley NRA, Millar K, Millica TA, Boyce BA, Antoniw P, Mountain A, Begent RHJ, Shochat D, Yarranton GT. Improved tumor targeting with chemically cross-linked recombinant antibody fragments.Cancer Res 1994; 54: 6176–6185.
    Google Scholar
  10. Pack P, Kujau M, Schroeckh U, Knüpfer R, Wenderoth R, Riesenberg D, Phickthun A. Improved bivalent miniantibodies with identical avidity as whole antibodies produced by high cell density fermentation of_Escherichia coli_.Bio Technology 1993; 11: 1271–1277.
    Google Scholar
  11. Marks JD, Griffiths AD, Malmqvist M, Clackson TP, Bye JM, Winter G. By-passing immunization: building high-affinity human antibodies by chain shuffling.Bio Technology 1992; 10: 779–783.
    Google Scholar
  12. Barbas CF III, Hu D, Dunlop N, Sawyer L, Cababa] D, Hendry RM, Nara PL, Burton DR. In vitro evolution of a neutralizing human antibody to human immunodeficiency virus type 1 to enhance affinity and broaden strain cross-reactivity.Proc Natl Acad Sci USA 1994; 91: 3809–3813.
    Google Scholar

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Authors and Affiliations

  1. Dipartimento di Medicina Sperimentale, Sezione di Medicina Nucleare, Policlinico Umberto I, Universita' di Roma “La Sapienza”, Via Regina Elena 324, I-00161, Roma, Italy
    Mauro Liberatore, Anna Paola Iurilli, Fabio Ponzo & Antonio Centi Colella
  2. Cambridge Centre for Protein Engineering — MRC Centre, Hills Road, CB2 2QH, Cambridge, UK
    Dario Neri
  3. Dipartimento di Biologia Molecolare, Universita' di Siena, Centro Didattico loc. Le Scotte, I-53100, Siena, Italy
    Giovanni Neri & Alessandro Pini
  4. Laboratorio di Biochimica degli Ormoni Sessuali, II Instituto di Clinica Ostetrica e Ginecologica, Universita' di Roma “La Sapienza”, Via Regina Elena 324, I-00161, Roma, Italy
    Giuseppe Spampinato, Fabrizio Padula & Alessandro Pala

Authors

  1. Mauro Liberatore
  2. Dario Neri
  3. Giovanni Neri
  4. Alessandro Pini
  5. Anna Paola Iurilli
  6. Fabio Ponzo
  7. Giuseppe Spampinato
  8. Fabrizio Padula
  9. Alessandro Pala
  10. Antonio Centi Colella

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Liberatore, M., Neri, D., Neri, G. et al. Efficient one-step direct labelling of recombinant antibodies with technetium-99m.Eur J Nucl Med 22, 1326–1329 (1995). https://doi.org/10.1007/BF00801622

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