title: Wide dynamic range of surface-plasmon-resonance-based assay for hepatitis B surface antigen antibody optimal detection in comparison with ELISA.
abstract: Limit of detection (LOD), limit of quantification, and the dynamic range of detection of hepatitis B surface antigen antibody (anti-HBs) using a surface plasmon resonance (SPR) chip-based approach with Pichia pastoris-derived recombinant hepatitis B surface antigen (HBsAg) as recognition element were established through the scouting for optimal conditions for the improvement of immobilization efficiency and in the use of optimal regeneration buffer. Recombinant HBsAg was immobilized onto the sensor surface of a CM5 chip at a concentration of 150 mg/L in sodium acetate buffer at pH 4 with added 0.6% Triton X-100. A regeneration solution of 20 mM HCl was optimally found to effectively unbind analytes from the ligand, thus allowing for multiple screening cycles. A dynamic range of detection of ∼0.00098-0.25 mg/L was obtained, and a sevenfold higher LOD, as well as a twofold increase in coefficient of variance of the replicated results, was shown as compared with enzyme-linked immunosorbent assay (ELISA). Evaluation of the assay for specificity showed no cross-reactivity with other antibodies tested. The ability of SPR chip-based assay and ELISA to detect anti-HBs in human serum was comparable, indicating that the SPR chip-based assay with its multiple screening capacity has greater advantage over ELISA.
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.|Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Yew Joon
authors: YJ
authors: Tam
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.|Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Nazariah Allaudin
authors: NA
authors: Zeenathul
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.|Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Morvarid Akhavan
authors: MA
authors: Rezaei
authors: Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Nor Hidayah
authors: NH
authors: Mustafa
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Mohd Lila Mohd
authors: MLM
authors: Azmi
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Abdul Rani
authors: AR
authors: Bahaman
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.|Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Sewn Cen
authors: SC
authors: Lo
authors: School of Industrial Technology, Universiti Sains Malaysia, Penang, Malaysia.
authors: Joo Shun
authors: JS
authors: Tan
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Homayoun
authors: H
authors: Hani
authors: Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia.
authors: Abdullah
authors: A
authors: Rasedee
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay; D006510:Hepatitis B Antibodies / Q000097:blood* / Q000378:metabolism*; D006514:Hepatitis B Surface Antigens / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism*; D057230:Limit of Detection; D016014:Linear Models; D046210:Microfluidic Analytical Techniques; D010843:Pichia; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D055767:Immobilized Proteins; D011994:Recombinant Proteins
keywords: Pichia pastoris; anti-HBs; chip-based detection assay; hepatitis B surface antigen; surface plasmon resonance