HUMAN SERUM FOLATE CAN BE MEASURED USING FOLATE BINDING PROTEIN LINKED TO ENZYME-LABELED PROTEIN LIGAND BINDING ASSAY (ELPLBA) AS WELL AS ELISA

Muhamad Arif Budiman, Mohamad Sadikin, Ani Retno Prijanti

Abstract


Background: Folate is an important substance used for purine and pyrimidine nucleotide synthesis. One measurement of folate that already establishes is using ELISA (Enzyme-linked immunosorbent assay) method. Folate binding protein is a protein that can bind folate, therefore it considered can be used as a tool that can replace antibody dependent ELISA method.

Objectives: The aim of this research was to create a method for folate measurement in serum called Enzyme-labeled protein ligand binding assay (ELPLBA) by replacing antibody as used in ELISA method with folate binding protein (FBP) that purified from the whey of milk.

Methods: The method is tested using 20 serum samples and compared to ELISA. Folate binding protein was purified from bovine’s milk using ammonium sulfate up to 90% saturated, DEAE-cellulose anion exchange chromatography and affinity chromatography. SDS-PAGE and western blot were used to establish the protein band of FBP that has molecular weight of ~25-35 kDa. ELPLBA was arranged with stationary phase using aminohexyl-agarose, and folic acid linked on it using carbodiimide.

Results: The result show there was no significant difference of folate concentration between ELPLBA (14.804 ± 2.795) and ELISA method (13.859 ± 3.638), p = 0.363.

Conclusion:  ELPLBA method show similarity for determination of folate in serum which was the same as standard folate measurement (ELISA).


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DOI: https://doi.org/10.32889/actabioina.v1i2.17

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