Immunological Separation of Bioactive Natural Compounds from Crude Drug Extract and Its Application for Cell-Based Studies

Abstract

In this study, we present a review on a useful approach, namely, immunoaffinity column coupled with monoclonal antibodies (MAbs), to separate natural compounds and its application for cell-based studies. The immunoaffinity column aids in separating the specific target compound from the crude extract. The column capacity was stable even after more than 10 purification cycles of use under the same conditions. After applying the crude extract to the column, the column was washed with washing buffer and eluted with elution buffer. The elution fraction contained the target compound bound to MAb, whereas the washing fraction was the crude extract, which contained all compounds except a group of target compounds; therefore, the washing fraction was referred to as a knockout (KO) crude extract. Cell-based studies using the KO extract revealed the actual effects of the natural compounds in the crude extract. One-step separation of natural compounds using the immunoaffinity column coupled with MAbs may help in determining the potential functions of natural compounds in crude extracts.

Simplified Column Fractionation with DPPH Assay Guidance: Isolated a Potent Hepatoprotective Fraction from Cassia auriculata

“…[24] Combining the benefits of online and offline screening methodologies, such as 1536 well micro-fractionation, can allow for fast fraction collection while maintaining high resolution achieved by the liquid chromatography separation. [25,26] Finally, bioactivity-directed fractionation and the testing of all column fractions for specific bioactivity can aid in the isolation of active compounds. [27,28] The present study aims to employ simplified column fractionation guided by DPPH assay to isolate a potent hepatoprotective fraction from Cassia auriculata extract.…”