Determination and Purification of Ricin Protein from Ricinus communis L. Seeds using CLC (Column Liquid Chramotagoraphy) and FPLC (Fast Protein Liquid Chramatography)

Abstract

Ricin is the most toxic substance isolated from Ricinus communis L. seeds. It is a heterodimeric two-domain polypeptide protein that includes chain A (30 kDa) and chain B (35 kDa) linked by a disulfide bond. Ricin binds to cells by the B chain and is then internalized. Ricin has been reported as a potential chemical for cancer treatment. The measurement of protein concentration in an aqueous sample is an important assay in biochemistry research. However, so far, the quantification of ricin protein is not much reported. In this study, the quantification of ricin protein extracted from R. communis L. from Nganjuk, East Java, Indonesia, the techniques purification was used column liquid chromatography (CLC) and fast protein liquid chromatography (FPLC), followed by quantification protein content using the Bradford method. Results showed that all purification techniques positively confirm the presence of ricin protein. Ricin protein content was 0.171±0.021 mg/mL and 0.382±0.023 mg/mL using CLC and FPLC, respectively. This study might contribute to understanding the chemical properties of the ricin protein from R. communis L. The Bradford method was used to quantify purified ricin, and the fractions obtained from FPLC were higher than those obtained from CLC.