Purification and characterization of polyclonal antibody against acrylamide

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Abstract

In our previous study, we have succeeded in preparing antibody against acrylamide using polyclonal method by immunization of N-acryloxysuccinimide conjugated with bovine serum albumin (NAS-BSA) into female New Zealand white rabbits. The aim of this study is to purify and characterize the produced antibody. The purification was performed using ammonium sulfate precipitation of IgG methods. Characterization was conducted in three series of serological tests including agar gel precipitation test (AGPT), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and dot blot immunoassay (DBIA). Concentration of the crudes and purified ammonium sulfate were found to be 20.876 and 8.056 ug/mL, respectively. The AGPT test of crude antibody showed a positive result against the antigen, while the SDS-PAGE showed that the purified ammonium sulfate antibody generated three molecular bands at 60, 50 and 25 kDa, suggested that the purified ammonium sulfate antibody was more specific than crude antibodies. In the case of DBIA test, crude and purified ammonium sulfate antibodies showed positive results against acrylamide. The results indicated that the NAS-BSA polyclonal antibody against acrylamide was successfully purified by using this method.

Original languageEnglish
Title of host publicationProceedings of the 3rd International Symposium on Current Progress in Mathematics and Sciences 2017, ISCPMS 2017
EditorsRatna Yuniati, Terry Mart, Ivandini T. Anggraningrum, Djoko Triyono, Kiki A. Sugeng
PublisherAmerican Institute of Physics Inc.
ISBN (Electronic)9780735417410
DOIs
Publication statusPublished - 22 Oct 2018
Event3rd International Symposium on Current Progress in Mathematics and Sciences 2017, ISCPMS 2017 - Bali, Indonesia
Duration: 26 Jul 201727 Jul 2017

Publication series

NameAIP Conference Proceedings
Volume2023
ISSN (Print)0094-243X
ISSN (Electronic)1551-7616

Conference

Conference3rd International Symposium on Current Progress in Mathematics and Sciences 2017, ISCPMS 2017
CountryIndonesia
CityBali
Period26/07/1727/07/17

Keywords

  • SDS-PAGE
  • ammonium sulfate precipitation
  • characterization
  • dot blot Immunoassay
  • purification

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    Assaat, L. D., Anggraningrum, I. T., & Saefudin, E. (2018). Purification and characterization of polyclonal antibody against acrylamide. In R. Yuniati, T. Mart, I. T. Anggraningrum, D. Triyono, & K. A. Sugeng (Eds.), Proceedings of the 3rd International Symposium on Current Progress in Mathematics and Sciences 2017, ISCPMS 2017 [020106] (AIP Conference Proceedings; Vol. 2023). American Institute of Physics Inc.. https://doi.org/10.1063/1.5064103