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Conjugation of proteins with chelating polymers via water-soluble carbodiimide andN-hydroxysulfosuccinimide

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Abstract

Water-soluble chelating polymers (CP) based on polylysine and diethylenetriaminepentaacetic acid (DTPA) have been prepared. The effect of the number of DTPA-groups in the polymer chain on the process of CP carbodiimide-mediated coupling to proteins has been studied. CP obtained were conjugated with proteins via carbodiimide andN-hydroxysulfosuccinimide (HSSI). The optimal conditions of CP activation were determined using model low-molecular-weight amine. It was shown that the addition of HSSI to an activation mixture increases the coupling efficiency of CP with immunoglobulins by 3‐4-fold compared with carbodiimide alone. Possible mechanisms of this phenomenon are discussed.

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Abbreviations

CP:

chelating polymer

DTPA:

diethylenetriaminepentaacetic acid

caDTPA:

cyclic anhydride of DTPA

HSSI:

N-hydroxusulfosuccinimide

EDC:

1-ethyl-3(3′-dimethylaminopropyl)-carbodiimide

TME:

tryptophan methyl ester

TNBS:

trinitrobenzene sulfonic acid

CT:

α-chymotrypsin

PL:

polylysine

ATEE:

N-acetyl-L-tyrosine ethyl ester.

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Authors and Affiliations

  1. USSR Cardiology Research Center, Institute of Experimental Cardiology, 3rd Cherepkovskaya St. 15A, 121552, Moscow, USSR

    A. L. Klibanov, M. A. Sllnkin & V. P. Torchilin

Authors
  1. A. L. Klibanov
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  2. M. A. Sllnkin
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  3. V. P. Torchilin
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Klibanov, A.L., Sllnkin, M.A. & Torchilin, V.P. Conjugation of proteins with chelating polymers via water-soluble carbodiimide andN-hydroxysulfosuccinimide. Appl Biochem Biotechnol 22, 45–58 (1989). https://doi.org/10.1007/BF02922696

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