Cysteine-containing fusion tag for site-specific conjugation of therapeutic and imaging agents to targeting proteins

Marina V. Backer, Zoia Levashova, Richard M Levenson, Francis G. Blankenberg, Joseph M. Backer

Research output: Chapter in Book/Report/Conference proceedingChapter

23 Citations (Scopus)

Abstract

Targeted delivery of therapeutic and imaging agents requires conjugation of a corresponding payload to a targeting peptide or protein. The ideal procedure should yield a uniform preparation of functionally active conjugates and be translatable for development of clinical products. We describe here our experience with site-specific protein modification via a novel cysteine-containing fusion tag (Cys-tag), which is a 15-amino-acid (aa) N-terminal fragment of human ribonuclease I with the R4C substitution. Several Cys-tagged proteins and peptides with different numbers of native cysteines were expressed and refolded into functionally active conformation, indicating that the tag does not interfere with the formation of internal disulfide bonds. We also describe standardized procedures for site-specific conjugation of very different payloads, such as functionalized lipids and liposomes, radionuclide chelators and radionuclides, fluorescent dyes, drug-derivatized dendrimers, scaffold proteins, biotin, and polyethyleneglycol to Cys-tagged peptides and proteins, as well as present examples of functional activity of targeted conjugates in vitro and in vivo. We expect that Cys-tag would provide new opportunities for development of targeted therapeutic and imaging agents for research and clinical use.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages275-294
Number of pages20
Volume494
ISBN (Print)9781588299901
DOIs
StatePublished - 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume494
ISSN (Print)10643745

Fingerprint

Protein Transport
Cysteine
Proteins
Radioisotopes
Peptides
Therapeutics
Dendrimers
Pancreatic Ribonuclease
Chelating Agents
Biotin
Fluorescent Dyes
Liposomes
Disulfides
Lipids
Amino Acids
Research
Pharmaceutical Preparations

Keywords

  • Cysteine-containing tag
  • Dendrimers
  • Liposomes
  • Site-specific conjugation
  • Targeted drug delivery
  • Targeted imaging

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Backer, M. V., Levashova, Z., Levenson, R. M., Blankenberg, F. G., & Backer, J. M. (2008). Cysteine-containing fusion tag for site-specific conjugation of therapeutic and imaging agents to targeting proteins. In Methods in Molecular Biology (Vol. 494, pp. 275-294). (Methods in Molecular Biology; Vol. 494). Humana Press. https://doi.org/10.1007/978-1-59745-419-3_16

Cysteine-containing fusion tag for site-specific conjugation of therapeutic and imaging agents to targeting proteins. / Backer, Marina V.; Levashova, Zoia; Levenson, Richard M; Blankenberg, Francis G.; Backer, Joseph M.

Methods in Molecular Biology. Vol. 494 Humana Press, 2008. p. 275-294 (Methods in Molecular Biology; Vol. 494).

Research output: Chapter in Book/Report/Conference proceedingChapter

Backer, MV, Levashova, Z, Levenson, RM, Blankenberg, FG & Backer, JM 2008, Cysteine-containing fusion tag for site-specific conjugation of therapeutic and imaging agents to targeting proteins. in Methods in Molecular Biology. vol. 494, Methods in Molecular Biology, vol. 494, Humana Press, pp. 275-294. https://doi.org/10.1007/978-1-59745-419-3_16
Backer MV, Levashova Z, Levenson RM, Blankenberg FG, Backer JM. Cysteine-containing fusion tag for site-specific conjugation of therapeutic and imaging agents to targeting proteins. In Methods in Molecular Biology. Vol. 494. Humana Press. 2008. p. 275-294. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-59745-419-3_16
Backer, Marina V. ; Levashova, Zoia ; Levenson, Richard M ; Blankenberg, Francis G. ; Backer, Joseph M. / Cysteine-containing fusion tag for site-specific conjugation of therapeutic and imaging agents to targeting proteins. Methods in Molecular Biology. Vol. 494 Humana Press, 2008. pp. 275-294 (Methods in Molecular Biology).
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