Abstract
Aim:Meat authentication gives significance values in view of religious, food safety, public health, quality assurance, and legal concern. Most of the meat authentication is based on molecular assay; a simpler method to authenticate meat is needed to develop. An immunoassays technique may offer a solution for simpler test. The aim of our current study was to develop a polyclonal antibody of Sus scrofa vittatus (Sumateran wild boar) as an immunodiagnostic reagent candidate.Materials and Methods:Three male New Zealand white rabbits were used in this study for antibody production. Antigen used was meat extract of Sumateran wild boar, each rabbit was immunized with meat extract antigen (0.5 mg/ml) emulsified in Freund’s complete adjuvant at a 1:1 (v/v) ratio as much as 1 ml at subcutaneous route. Booster was carried out 3 times with interval time of 14 days, using meat extract antigen emulsified in Freund’s incomplete adjuvant at a 1:1 (v/v) ratio. Serum samples were taken every week, start from 1 week after the first immunization up to 1 week after the third booster. Antibody purification was performed using ammonium sulfate precipitation and Protein A. The presence of specific antibody was determined using agar gel precipitation test and enzyme-linked immunosorbent assay, while purified specific IgG was characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis method.Results:Specific antibody was detected at 14 days after the first immunization and still detected until 2 weeks after the third booster. Highest absorbance of specific antibody was detected 1 week after the third booster.Conclusion:The present study demonstrated that specific antibody of Sumateran wild boar is favorable to be produced in rabbit and showed that antibody produced is applicable to detect Sumateran wild boar meat antigen in immunodiffusion assay, indicating that it is promising as a reagent candidate in immunodiagnostic assay/kit.
Highlights
Consumption of meat and its product is increasing each year worldwide; adulteration of meat has been a long time issue
The present study demonstrated that specific antibody of Sumateran wild boar is favorable to be produced in rabbit and showed that antibody produced is applicable to detect Sumateran wild boar meat antigen in immunodiffusion assay, indicating that it is promising as a reagent candidate in immunodiagnostic assay/kit
agar gel precipitation test (AGPT) was conducted to confirm that the antibodies produced are able to bind with Sumateran wild boar meat antigen
Summary
Consumption of meat and its product is increasing each year worldwide; adulteration of meat has been a long time issue. In China, “hang a sheep head, sell vinegar” is a widely known proverb. Meat adulteration affects food safety, quality, and many other aspects and has becoming a public focus [1]. To overcome this problem, various analytical methods based on physical, chemical, anatomical, histological, and biological approaches are being utilized to identify meat species [2]. Sus scrofa vittatus (Sumateran wild boars) are superabundant in Sumateran forest [3], this condition has led to exploitation for commercial purpose [4].
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