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
Journal article Acta Biochimica Indonesiana • 2018

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(English, 9 pages)


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).




Acta Biochimica Indonesiana

Acta Biochimica Indonesiana (ActaBiolna) is an open access, peer-reviewed journal published by th... see more