Synthesis and characterization of polysialic acid-uricase conjugates for the treatment of hyperuricemia

Ardra Punnappuzha, Jagadeeshbabu Ponnanettiyappan, Reval Sekhar Nishith, Sanjay Hadigal, Preethi Ganapathi Pai

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Uricase, an enzyme used for the treatment of hyperuricemia, is conjugated with polysialic acid (PSA) of average molecular weight of 10 kDa by reductive amination in presence of NaCNBH3 in order to improve its pharmacological properties. Polysialylation with 50-,100-,150- and 200-fold molar excess of PSA increased the percentage substitution of the free amino groups on enzyme surface (46, 66, 78 and 80 % respectively). The SDS-PAGE is used to visualize the conjugates with increased molecular weight and it retained almost 65 % of their initial specific activity after conjugation. The stability studies at physiological condition reveals improved stability and activity than the native enzyme. The apparent KM of the enzyme has increased slightly from 4.18 × 10-5 M to 5.46 × 10-5 M suggesting that the affinity of the substrate to the enzyme has not been altered to a higher extent. The conjugates, when probed against anti-uricase antibodies generated in rabbit, showed a clean decline in the affinity by 35 % and also have retained double the catalytic activity than that of the native enzyme after exposure to antiserum. The results suggest that uricase-PSA conjugates can be used as an alternative to the conventional synthetic polymer-enzyme conjugates.

Original languageEnglish
Pages (from-to)465-472
Number of pages8
JournalInternational Journal of Peptide Research and Therapeutics
Volume20
Issue number4
DOIs
Publication statusPublished - 01-01-2014

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Urate Oxidase
Hyperuricemia
Enzymes
Acids
Molecular Weight
Molecular weight
Amination
polysialic acid
Antibodies
Immune Sera
Polyacrylamide Gel Electrophoresis
Anti-Idiotypic Antibodies
Catalyst activity
Polymers
Substitution reactions
Pharmacology
Rabbits
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Bioengineering
  • Molecular Medicine
  • Drug Discovery
  • Analytical Chemistry

Cite this

Punnappuzha, Ardra ; Ponnanettiyappan, Jagadeeshbabu ; Nishith, Reval Sekhar ; Hadigal, Sanjay ; Pai, Preethi Ganapathi. / Synthesis and characterization of polysialic acid-uricase conjugates for the treatment of hyperuricemia. In: International Journal of Peptide Research and Therapeutics. 2014 ; Vol. 20, No. 4. pp. 465-472.
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Synthesis and characterization of polysialic acid-uricase conjugates for the treatment of hyperuricemia. / Punnappuzha, Ardra; Ponnanettiyappan, Jagadeeshbabu; Nishith, Reval Sekhar; Hadigal, Sanjay; Pai, Preethi Ganapathi.

In: International Journal of Peptide Research and Therapeutics, Vol. 20, No. 4, 01.01.2014, p. 465-472.

Research output: Contribution to journalArticle

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