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1044 Kipriyanov<br />

nant antibodies stabilized by albumin can be stored at –80°C for years without loss of<br />

activity. These preparations may be used for a number of biological assays such as ELISA,<br />

flow cytometry, and analyses of antitumor activity both in vitro and in vivo.<br />

22. If different epitopes are used for immunodetection (non c-myc), the corresponding antibodies<br />

should be used.<br />

References<br />

1. Bird, R. E., Hardman, K. D., Jacobson, J. W., Johnson, S., Kaufman, B. M., Lee, S. M.,<br />

et al. (1988) Single-chain antigen-binding proteins. Science 242, 423–426.<br />

2. Huston, J. S., Levinson, D., Mudgett Hunter, M., Tai, M. S., Novotny, J., Margolies, M.<br />

N., et al. (1988) <strong>Protein</strong> engineering of antibody binding sites: recovery of specific activity<br />

in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc. Natl. Acad.<br />

Sci. USA 85, 5879–5883.<br />

3. Milenic, D. E., Yokota, T., Filpula, D. R., Finkelman, M. A., Dodd, S. W., Wood, J. F.,<br />

et al. (1991) Construction, binding properties, metabolism, and tumor targeting of a singlechain<br />

Fv derived from the pancarcinoma monoclonal antibody CC49. Cancer Res. 51,<br />

6363–6371.<br />

4. Yokota, T., Milenic, D. E., Whitlow, M., and Schlom, J. (1992) Rapid tumor penetration of<br />

a single-chain Fv and comparison with other immunoglobulin forms. Cancer Res. 52,<br />

3402–3408.<br />

5. Yokota, T., Milenic, D. E., Whitlow, M., Wood, J. F., Hubert, S. L., and Schlom, J. (1993)<br />

Microautoradiographic analysis of the normal organ distribution of radioiodinated singlechain<br />

Fv and other immunoglobulin forms. Cancer Res. 53, 3776–3783.<br />

6. Adams, G. P., McCartney, J. E., Tai, M. S., Oppermann, H., Huston, J. S., Stafford, W. F.,<br />

et al. (1993) Highly specific in vivo tumor targeting by monovalent and divalent forms of<br />

741F8 anti-c-erbB-2 single-chain Fv. Cancer Res. 53, 4026–4034.<br />

7. Gruber, M., Schodin, B. A., Wilson, E. R., and Kranz, D. M. (1994) Efficient tumor cell<br />

lysis mediated by a bispecific single-chain antibody expressed in Escherichia coli.<br />

J. Immunol. 152, 5368–5374.<br />

8. Kurucz, I., Titus, J. A., Jost, C. R., Jacobus, C. M., and Segal, D. M. (1995) Retargeting of<br />

CTL by an efficiently refolded bispecific single-chain Fv dimer produced in bacteria.<br />

J. Immunol. 154, 4576–4582.<br />

9. Kipriyanov, S. M., Moldenhauer, G., Schuhmacher, J., Cochlovius, B., Von der Lieth, C. W.,<br />

Matys, E. R., and Little, M. (1999) Bispecific tandem diabody for tumor therapy with<br />

improved antigen binding and pharmacokinetics. J. Mol. Biol. 293, 41–56.<br />

10. Kontermann, R. E. and Müller, R. (1999) Intracellular and cell surface displayed singlechain<br />

diabodies. J. Immunol. Meth. 226, 179–188.<br />

11. Kipriyanov, S. M. and Little, M. (1999) Generation of recombinant antibodies. Mol.<br />

Biotechnol. 12, 173–201.<br />

12. Pugsley, A. P. (1993) The complete general secretory pathway in gram-negative bacteria.<br />

Microbiol. Rev. 57, 50–108.<br />

13. Whitlow, M. and Filpula, D. (1991) Single-chain Fv proteins and their fusion proteins.<br />

Meth. Comp. Meth. Enzymol. 2, 97–105.<br />

14. Kipriyanov, S. M., Dübel, S., Breitling, F., Kontermann, R. E., and Little, M. (1994)<br />

Recombinant single-chain Fv fragments carrying C-terminal cysteine residues: production<br />

of bivalent and biotinylated miniantibodies. Mol. Immunol. 31, 1047–1058.<br />

15. Plückthun, A. (1994) Antibodies from Escherichia coli, in Handbook of Experimental<br />

Pharmacology, Vol. 113: The Pharmacology of Monoclonal Antibodies (Rosenberg, M.<br />

and Moore, G. P., eds.), Springer-Verlag, Berlin, Heidelberg, pp. 269–315.

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