The functional effects of biotinylation of anti-angiotensin-converting enzyme monoclonal antibody in terms of targeting in vivo

Vladimir R. Muzykantov, Vitaly D. Gavriluk, Alexander Reinecke, Elena Nikolaevna Atochina, Alice Kuo, Elliot S. Barnathan, Aron B. Fisher

Результат исследований: Материалы для журналаСтатьярецензирование

22 Цитирования (Scopus)

Аннотация

The effect of modification with biotin N-hydroxysuccinimide ester of mouse monoclonal antibody to angiotensin-converting enzyme, anti-ACE Mab 9B9, on its targeting to endothelial cells has been studied in vitro and in vivo. By in vitro assay, Mab 9B9 biotinylated at a biotin/IgG molar ratio in reaction mixture (B/IgG ratio) of 0.7-2.2 bound streptavidin monovalently and retained antigen-binding capacity. Mab 9B9 biotinylated at a B/IgG ratio of 20 and higher bound streptavidin polyvalently. Extensive biotinylation (B/IgG ratio of 60 and higher) led to dramatic reduction of Mab 9B9 Ag-binding capacity and to reduction of Mab 9B9 recognition by goat polyclonal antibody to mouse IgG. Radiolabeled Mab 9B9 biotinylated at a B/IgG ratio of 6 (b6-Mab 9B9) bound effectively to cultured vascular endothelium, with affinity characteristics similar to non-biotinylated Mab 9B9. Endothelial cells internalized both Mab 9B9 and b6-Mab 9B9 to the same extent (60% internalization at 3 h incubation at 37°C). Degradation of cell surface-associated Mab 9B9 or b6-Mab 9B9 was very low (<1% as measured by TCA solubility of radiolabel). In contrast, degradation of internalized b6-Mab 9B9 was more profound than that of Mab 9B9 (20 ± 3% vs 6 ± 1%, P <0.01). After injection in rats, radiolabeled b6-Mab 9B9 had a biodistribution pattern similar to that of radiolabeled Mab 9B9. Both preparations effectively accumulated in the lung (15-20% of injected dose/g of tissue vs 2% of injected dose/g of blood). Extensive biotinylation led to both a reduction of specific pulmonary uptake of Mab 9B9 and an enhanced blood clearance of Mab 9B9. Streptavidin binding to b6-Mab 9B9 did not alter biodistribution or pulmonary targeting of biotinylated antibody. We conclude that extensive biotinylation induces complex alterations of the functional properties of Mab 9B9. In contrast, b6-Mab 9B9 may serve as an affinity carrier for targeting of biotinylated compounds to the pulmonary endothelium.

Язык оригиналаАнглийский
Страницы (с-по)279-287
Число страниц9
ЖурналAnalytical Biochemistry
Том226
Номер выпуска2
DOI
СостояниеОпубликовано - 1995
Опубликовано для внешнего пользованияДа

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

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