Synthesis of Polyclonal Antibodies against Aflatoxin B1

Wicaksono, Wiyogo Prio; Permana, Deni Samsudin; Maryam, Romsyah; Einaga, Yasuaki

doi:10.7454/mss.v19i3.4847

Cited by 3 publications

(6 citation statements)

References 13 publications

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“…The AFB1-CMO hapten was generated according to the method of Wicaksono with some modifications [17]. Briefly, 60 µl of a mixture of methanol/pyridine/water (4:1:1) containing 16 µg of AFB1 and 16.8 µg of CMO was refluxed at 90 o C in the dark for 2 hours, followed by evaporation at 65 o C, 150 mbar until dry to remove pyridine.…”

Section: Synthesis Of the Aflatoxin B1-carboxymethyl Hydroxylamine Hementioning

confidence: 99%

“…3A, lane 3-6). This reaction was carried out at 86 o C by Wicaksono et al [17] and at 110 o C by Kim et al [18]. We observed that the spot intensity of AFB1-CMO was decreased at 110 o C. Therefore, suitable temperature for coupling reaction of AFB1 and CMO should be in a range of 80 -100 o C. Four different reaction times of 1, 2, 3 and 4 hours were investigated to identify the suitable reaction time for coupling AFB1 and CMO.…”

Section: Optimizing the Reaction Temperatures And Timesmentioning

confidence: 99%

“…3B). This reaction was carried out for 3 hours by Khademi et al [19] and even for 5 hours by Wicaksono et al [17].…”

Section: Optimizing the Reaction Temperatures And Timesmentioning

confidence: 99%

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Efficient Conjugation of Aflatoxin B1 With Bovine Serum Albumin Applying for Development of Aflatoxin B1 Quick Test

Phong

Phạm²,

Nguyen

et al. 2018

JST

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Aflatoxins are secondary metabolites mostly produced by Aspergillus flavus and Aspergillus parasiticus and found in agricultural foodstuff such as maize grains, peanuts, animal feeds,... These are toxic and cancerous agents for humans and animals. Among them, aflatoxin B1 is the most consideration due to its highest toxicity and presence in samples. Several methods were developed to detect aflatoxin in food, feed and other foodstuffs such as chromatographic methods (TLC, HPTLC, HPLC), LC-MS/MS, FTIR, RIA, ELISA, SPR, electrochemical, and immunodipstick. Among them only immunodipstick method is compatible for field usage. Therefore, aim of the present study is to determine optimal condition for conjugation of aflatoxin B1 (AFB1) with bovine serum albumin (BSA) to develop a lateral flow immunoassay test strip for detection of aflatoxin B1. Optimal conditions for generating mediate compound of aflatoxin B1 – CMO was investigated: AFB1:CMO ratio of 1:2, reflux temperature of 100oC for 2 hours. Concentration of aflatoxin B1 in reaction was 0.27 µg/µl. Conjugation of AFB1 with CMO was confirmed by thin layer chromatography, HPLC and FT-IR methods. Furthermore, optimal conditions for conjugation of mediate AFB1-CMO derivative with BSA were also investigated. Ratio of AFB1-CMO with BSA was determined as 40:1 and equal to predicted theoretical ratio. Efficient conjugation conditions were 25oC for 2 hours in bicarbonate buffer pH 9.5. Generated conjugate of AFB1-BSA was successfully applied to construct a lateral flow immunoassay test strip for detection of aflatoxin B1

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Section: Synthesis Of the Aflatoxin B1-carboxymethyl Hydroxylamine Hementioning

confidence: 99%

Section: Optimizing the Reaction Temperatures And Timesmentioning

confidence: 99%

See 1 more Smart Citation

Efficient Conjugation of Aflatoxin B1 With Bovine Serum Albumin Applying for Development of Aflatoxin B1 Quick Test

Phong

Phạm²,

Nguyen

et al. 2018

JST

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“…The development of such methods includes a method of a specific antibody application, which can be prepared by immunizing animals such as rabbits, mice, goats, hamsters, and chickens with an immunogen [5]. Wicaksono et al has produced aflatoxin antibody by administering immunogen (aflatoxin B1-carboxymethyl hydroxylamine hemihydrochloride-BSA) into rabbits [3]. This study was performed for further purification of the antibody.…”

Section: Introductionmentioning

confidence: 99%

“…Aflatoxin testing methods such as by HPLC, TLC, and ELISA have been reported [3][4][5]. The development of such methods includes a method of a specific antibody application, which can be prepared by immunizing animals such as rabbits, mice, goats, hamsters, and chickens with an immunogen [5].…”

Section: Introductionmentioning

confidence: 99%

Purification of aflatoxin B1 antibody for the development of aflatoxin biosensor

Prihantoro

Saepudin²,

Ivandini³

2017

AIP Conference Proceedings

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Production of Newcastle Disease Polyclonal Antibody as the Alternative of Immunohistochemistry Primary Antibody against Newcastle Disease in Poultry

Naf’an¹,

Kurniasih²,

Untari³

et al. 2021

WVJ

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Newcastle disease (ND) is the most pathogenic viral infection in poultry. Furthermore, the availability of laboratories that support the molecular diagnosis of ND is still limited in Indonesia. The present study aimed to produce ND polyclonal antibody as the alternative of immunohistochemistry primary antibody against ND in poultry. Two adult male New Zealand White rabbits weighed 2.5 kg were vaccinated seven days after the adaptation using intraperitoneal injection of the ND live vaccine at multilevel doses weekly. The serum was collected inactivated, and purified in the sixth week. A total number of 31 chicken samples were collected and their samples of brain, lung, spleen, and intestine were tested using immunohistochemistry and Reverse Transcription Polymerase Chain reaction (RT-PCR). The result showed that 19/31 (61%) were positive against immunohistochemistry and RT-PCR and a total of 12/31 (39%) were negative. Based on the obtained results, immunohistochemistry using ND polyclonal antibody had a similar accuracy with RT-PCR. It can be concluded that ND polyclonal antibody produced by vaccination in the rabbit could be used as the alternative immunohistochemistry primary antibody for diagnosing ND in poultry.

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Synthesis of Polyclonal Antibodies against Aflatoxin B1

Cited by 3 publications

References 13 publications

Efficient Conjugation of Aflatoxin B1 With Bovine Serum Albumin Applying for Development of Aflatoxin B1 Quick Test

Efficient Conjugation of Aflatoxin B1 With Bovine Serum Albumin Applying for Development of Aflatoxin B1 Quick Test

Purification of aflatoxin B1 antibody for the development of aflatoxin biosensor

Production of Newcastle Disease Polyclonal Antibody as the Alternative of Immunohistochemistry Primary Antibody against Newcastle Disease in Poultry

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