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All Journal 3BIO: Journal of Biological Science, Technology and Management Current Research on Biosciences and Biotechnology
Niloperbowo, Wardono
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience

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Anti-HBsAg IgY polyclonal antibodies potential as capture antibody for HBsAg detection kit development A'yun, Ramadhani Qurrota; Hakim, Meutia Diva; Giri-Rachman , Ernawati Arifin; Tan, Marselina; Niloperbowo, Wardono
Current Research on Biosciences and Biotechnology Vol. 5 No. 2 (2024)
Publisher : Institut Teknologi Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5614/crbb.2024.5.2/UBVBMKPZ

Abstract

Hepatitis B, affecting about 296 million globally, is a significant concern, with Indonesia ranking second in Southeast Asia for case numbers. The disease's latent initial phase, devoid of early symptoms upon hepatitis B virus (HBV) infection, highlights the demand for precise diagnostics. This research aimed to develop anti-HBsAg polyclonal antibodies (pAb) for application as capture agents within a sandwich enzyme-linked immunosorbent assay (ELISA). Chicken egg-derived IgY antibodies have advantages over mammalian ones due to simpler extraction and higher yield. In a study involving 21-week-old chickens, four intramuscular injections of 500 µg HBsAg antigen in Freund's adjuvant were administered at two-week intervals. Eggs as IgY source were collected daily and then IgY was isolated from eggyolk using polyethylene glycol (PEG) precipitation. The Bradford method was used to measure the total protein concentration, while the existence of IgY and pAb specific IgY against Anti-HBsAg was verified through SDS-PAGE and sandwich ELISA using HRP as a reporter, respectively. The resulting SDS-PAGE showed two distinct IgY bands: a 68 kDa heavy chain and a 23 kDa light chain. Using these anti-HBsAg IgY antibodies as capturing agents, the slightly elevation of IgY pAb against HBsAg level has been identified within the second week following the initial immunization. Subsequently, from the third to the eighth week, antibody levels escalated significantly, ranging from 2 to 13-fold higher than those observed in the second week. These findings suggest the potential use of IgY pAb as effective capture antibodies in sandwich ELISA for HBsAg antigen detection.
The Influence of Polyethylene Glycol Precipitation Methods on Yield and Purity of White Radish Peroxidase Faizah, Nur Al; Giri-Rachman, Ernawati Arifin; Niloperbowo, Wardono
3BIO: Journal of Biological Science, Technology and Management Vol. 6 No. 2 (2024)
Publisher : School of Life Sciences and Technology, Institut Teknologi Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5614/3bio.2024.6.2.5

Abstract

Proteins are widely used in various industries as highly valued biotechnology products. One example is horseradish peroxidase isolated from horseradish (Armoracia rusticana) that used as enzyme label in immunochemistry. However, the cultivation of horseradish is limited to subtropical countries, making the dependency on horseradish peroxidase unsustainable for tropical countries. Numerous studies have explored alternative peroxidases, and white radish peroxidase isolated from Raphanus sativus L. has emerged as a promising candidate. In this study, white radish peroxidase is isolated using the polyethylene glycol (PEG) precipitation method which is widely used as a simple and cost-effective method. This study aims to evaluate the effectiveness of the one-step and two-step PEG precipitation method. The one-step PEG precipitation method used in this study was done by mixing the white radish juice with PEG 6000 30% (w/v), while the two-step method was done by mixing it with PEG 400 20% (w/v) and PEG 6000 30% (w/v) consecutively. This study compares the yield and recovery levels of total protein and white radish peroxidase, as well as the enzymatic specific activity of white radish peroxidase isolated both by the one-step PEG precipitation and the two-step PEG precipitation. The results indicate that both extraction methods yield the same level of white radish peroxidase. However, they differ in terms of purity. The two-step extraction method results in white radish peroxidase with higher purity, as evidenced by its specific activity towards the chromogen ABTS in the presence of H2O2.
Bioconversion of inorganic selenium to organic selenium in the black soldier fly (BSF) larvae Sihombing, Rolina Anna Erica; Niloperbowo, Wardono; Nugrahapraja, Husna
Current Research on Biosciences and Biotechnology Vol. 7 No. 1 (2025)
Publisher : Institut Teknologi Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5614/crbb.2025.7.1/JAFFPU8L

Abstract

Selenium is one of the essential micronutrients needed to fulfil livestock nutrition, which can be found in inorganic and organic forms. Black Soldier Fly (BSF) larvae can potentially be used as a natural converter from inorganic selenium to organic selenium. However, the capacity and response of BSF larvae to convert selenium are still unknown. This study aims to determine the effect of inorganic selenium administration on BSF larvae. The research method was the determination of selenium concentration by UV-Vis spectrophotometry method based on the variation in the age of inorganic selenium administration in BSF larvae to the growth of BSF larvae, and the accumulation of selenium in BSF larvae, with variations in age of 0, 4, 8, and 12 days of age given sodium selenite (Na2SeO3) and control without administration of sodium selenite from the beginning to the end of rearing. There was no significant difference (p > 0.05) in the growth performance of BSF larvae and the accumulation of selenium in BSF larvae in the age variation experiment of BSF larvae when given sodium selenite. Based on the acquisition of larval mass, the growth rate of BSF larvae in the control treatment, with sodium selenite at 1 mg/kg, was 0.129, 0.093, 0.037, 0.156, and 0.128 mg/day at 0, 4, 8, and 12 days, respectively. These results indicate that inorganic selenium administration to BSF larvae can occur during the rearing period. In the experiment of variation in the concentration of inorganic selenium given to BSF larvae, the growth rate of BSF larvae was significantly higher (p < 0.05) when given 1000 mg/kg of sodium selenite, which was 0.467 mg/day.
Co-Authors A'yun, Ramadhani Qurrota ERNAWATI ARIFIN GIRI-RACHMAN Faizah, Nur Al Giri-Rachman , Ernawati Arifin Hakim, Meutia Diva Marselina Tan Nugrahapraja, Husna Sihombing, Rolina Anna Erica