| Prolonged circulating lives of single-chain Fv proteins conjugated with polyethylene glycol: a comparison of conjugation chemistries and compounds. | |
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MedLine Citation:
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PMID: 10563766 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The utility of single-chain Fv proteins as therapeutic agents would be substantially broadened if the circulating lives of these minimal antigen-binding polypeptides were both prolonged and adjustable. Poly(ethylene glycol) (PEG) bioconjugate derivatives of the model single-chain Fv, CC49/218 sFv, were constructed using six different linker chemistries that selectively conjugate either primary amines or carboxylic acid groups. Activated PEG polymers with molecular weights of 2000, 5000, 10 000, 12 000, and 20 000 were included in the sFv bioconjugate evaluation. Additionally, the influence of PEG conjugate geometry in branched PEG strands (U-PEG) and the effect of multimeric PEG-sFv bioconjugates on circulating life and affinity were examined. Although random and extensive PEG polymer conjugations have been achievable in highly active derivatives of the prototypical PEG-enzymes, PEGylation of CC49/218 sFv required stringent adjustment of reaction conditions in order to preserve antigen-binding affinity as measured in either mucin-specific or whole cell immunoassays. Purified bioconjugates with PEG:sFv ratios of 1:1 through 2:1 were identified as promising candidates which exhibit sFv affinity (K(d)) values within 2-fold of the unmodified sFv protein. Interestingly, PEG conjugation to carboxylic acid moieties, using a PEG-hydrazide chemistry, achieved significant activity retention in bioconjugates at a higher PEG:sFv ratio (5:1) than with any of the amine-reactive activated PEG polymers. Prolonged circulating life in mice was demonstrated for each of the PEG conjugates. An increase in PEG polymer length was found to be more effective for serum half-life extension than a corresponding increase in total PEG mass. For example, CC49/218 sFv conjugated to either one strand of PEG-20000, or four strands of PEG-5000, displayed about 20- or 14-fold increased serum half-life, respectively, relative to the unmodified sFv. The demonstrated suitability of established random conjugation chemistries for PEGylation of sFv proteins, in conjunction with innovative site-specific conjugation methods, indicates that production of a panoply of sFv proteins with both engineered affinity and tailored circulating life may now be achievable. |
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Authors:
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L S Lee; C Conover; C Shi; M Whitlow; D Filpula |
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Publication Detail:
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Type: Comparative Study; Journal Article |
Journal Detail:
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Title: Bioconjugate chemistry Volume: 10 ISSN: 1043-1802 ISO Abbreviation: Bioconjug. Chem. Publication Date: 1999 Nov-Dec |
Date Detail:
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Created Date: 2000-01-06 Completed Date: 2000-01-06 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 9010319 Medline TA: Bioconjug Chem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 973-81 Citation Subset: IM |
Affiliation:
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Enzon Inc./SCA Ventures, 20 Kingsbridge Road, Piscataway, New Jersey 08854-3969, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Antibodies, Monoclonal Antibodies, Neoplasm / blood, chemistry* Aspartic Acid / chemistry Chromatography, High Pressure Liquid Crystallization Electrophoresis, Polyacrylamide Gel Glutamic Acid / chemistry Glycosylation Immunoglobulin Fragments / blood*, chemistry* Kinetics Lysine / chemistry Models, Molecular Molecular Sequence Data Molecular Weight Polyethylene Glycols / chemistry* Structure-Activity Relationship |
| Chemical | |
Reg. No./Substance:
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0/Antibodies, Monoclonal; 0/Antibodies, Neoplasm; 0/B72.3 antibody; 0/Immunoglobulin Fragments; 0/Polyethylene Glycols; 0/immunoglobulin Fv; 56-84-8/Aspartic Acid; 56-86-0/Glutamic Acid; 56-87-1/Lysine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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