Comparative efficiency of ammonium sulfate alone and ammonium sulfate-octanoic acid combined methods in the purification potential of antibodies

Ahsan, Naveed

doi:10.19045/bspab.2019.80113

Cited by 4 publications

(4 citation statements)

References 12 publications

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“…The purification of pAbs was done by the combined AS-OA precipitation method described in ref ( 28 ) with some modifications. Serum (5 mL) was taken and diluted with an acetate buffer in a 1:3 ratio.…”

Section: Methodsmentioning

confidence: 99%

Development of a Lateral-Flow Immunochromatographic Strip for the Detection of Oxytetracycline Residues in Biological Fluids

Aleem,

Akhtar,

Muhammad

et al. 2023

ACS Omega

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Oxytetracycline (OTC) is extensively used in veterinary medicine and for growth promotion around the globe. The indiscriminate use of OTC in food-producing animals leaves residues in animal products. The presence of these residues in animal products causes economic losses and harmful effects on consumers. Different regulatory bodies set maximum residue limits (MRLs) for different tetracyclines. To avoid harmful effects, there is a need for a simple, fast, and economical method for the screening of animal products. In this study, a fast, economical, and user-friendly lateral-flow immunochromatographic (LFIC) assay based on gold nanoparticles (AuNPs) was developed to detect the presence of OTC residues in biological fluids. AuNPs provided visual results as red lines in 6−15 min. Polyclonal rabbit IgG antibodies were produced using the immunogen of OTC. These antibodies were purified by the combined ammonium sulfate-octanoic acid precipitation method. Antibodies were conjugated to AuNPs as recognition biomolecules. A LFIC strip was optimized using borate buffer spiked with different concentrations of the OTC. The visual limit of detection (LOD) in different biological samples (milk, serum, and urine) was determined using samples spiked with OTC. The LOD was found to be 15 μg/L, which is very low from the MRL (100 μg/L) set by different regulatory authorities. This LFIC strip can be used to detect OTC residues in biological fluids for point-of-care testing (POCT). These strips are easy to use, cost-effective, and portable and provide quick results without the use of laboratory instruments.

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Section: Methodsmentioning

confidence: 99%

Development of a Lateral-Flow Immunochromatographic Strip for the Detection of Oxytetracycline Residues in Biological Fluids

Aleem,

Akhtar,

Muhammad

et al. 2023

ACS Omega

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“…Anti‐ZEN monoclonal antibody was purified by ammonium sulfate and caprylic acid precipitation method (Naveed, 2019). QDs were conjugated with the anti‐ZEN mAb through the EDC crosslinking reaction to form fluorescent probes (Liu et al, 2021).…”

Section: Methodsmentioning

confidence: 99%

Fluorescence immunoassay based on phage mimotope for nontoxic detection of Zearalenone in maize

Zhou

Wang

et al. 2022

Journal of Food Safety

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Zearalenone (ZEN) is one of the most common mycotoxin contaminants worldwide. In this study, a phage-based direct competitive fluorescence immunosorbent assay (P-dcFLISA) was developed for the detection of ZEN. In this P-dcFLISA, phage mimotope was used to replace chemically synthesized antigens to improve the safety of experiments. Furthermore, ZnCdSe/ZnS (core/shell) quantum dots (QDs) as fluorescent labeling material replaced chromogenic substrate, which are used in traditional ELISA. The 50% inhibition value (IC 50 ) of P-dcFLISA was 0.301 ng/ml, which was approximately 12-fold lower than that of phage-based indirect competitive enzyme-linked immunosorbent assay (P-icELISA). The limit of detection (LOD) of P-dcFLISA was 0.023 ng/ml and the detection range was 0.060-1.531 ng/ml. In the added recovery assay, recovery rate was 93.18-105.52%, with the coefficient variation (CV) ranging from 8.39% to 10.55%. The establishment of P-icFLISA not only save time, but also avoid the use of mycotoxins in the detection process, providing a safe method for the detection of mycotoxins. In conclusion, phage-based fluorescence immunoassay method has great potential application in ZEN detection and agricultural monitoring.

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“…IgG monoclonal antibody from serum raised against mycotoxins was purified through the method of caprylic acid and ammonium sulfate precipitation. 36 As shown in Scheme 1B, the fluorescent probes were synthesized through the EDC conjugation method with a little modification. 31 In brief, 0.1 mg of the carboxyl groups of QBs-polyethylene glycol (PEG) (10 mg/mL) was activated by adding into 0.5 mL of 10 mM phosphate buffer (PB) solution (pH = 6.0) containing 1 mM EDC•HCl and 1 mM NHS.…”

Section: Methodsmentioning

confidence: 99%

“…IgG monoclonal antibody from serum raised against mycotoxins was purified through the method of caprylic acid and ammonium sulfate precipitation . As shown in Scheme B, the fluorescent probes were synthesized through the EDC conjugation method with a little modification .…”

Section: Methodsmentioning

confidence: 99%

Biosynthetic Mycotoxin Conjugate Mimetics-Mediated Green Strategy for Multiplex Mycotoxin Immunochromatographic Assay

Yan

You

et al. 2020

J. Agric. Food Chem.

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Various mycotoxins widely co-exist in agro-products, and their combined effects cause toxicity and potential carcinogenicity to humans and animals. In this work, we developed an economical and sensitive quantum dots (QDs)/QD microbead (QDs/QB)-based multiplex immunochromatographic assay (mICA) for the rapid detection of fumonisin B1 (FB1), zearalenone (ZEN), and ochratoxin A (OTA) without the building-up process of mycotoxin conjugates. QDs and QBs were selected as fluorescent reporters and conjugated with antimycotoxin monoclonal antibodies for improving sensitivity. Furthermore, phage-displayed FB1, ZEN, and OTA mimotope peptide-based soluble and monovalent fusions to maltose-binding protein (MBP) were applied onto the test line of the mICA as the mimetic coating antigen. Under the optimized conditions, the visual detection limits (vLODs) of peptide–MBP-based mICA could be obtained as 0.25 ng/mL for FB1, 3.0 ng/mL for ZEN, and 0.5 ng/mL for OTA within 10 min. The results for spiked real sample detection indicate good accuracy, reproducibility, and practicability. In addition, the proposed mICA was comparable with ultraperformance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) in terms of reliability in detecting FB1, ZEN, and OTA using natural samples. From the point of promoting commercial production, these time-saving and low-cost peptide–MBP antigens applied in ICA might provide promising potential for promoting productivity and decreasing the cost of production.

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Comparative efficiency of ammonium sulfate alone and ammonium sulfate-octanoic acid combined methods in the purification potential of antibodies

Abstract: Comparative efficiency of ammonium sulfate alone and ammonium sulfate-octanoic acid combined methods in the purification potential of antibodies.

Cited by 4 publications

References 12 publications

Development of a Lateral-Flow Immunochromatographic Strip for the Detection of Oxytetracycline Residues in Biological Fluids

Development of a Lateral-Flow Immunochromatographic Strip for the Detection of Oxytetracycline Residues in Biological Fluids

Fluorescence immunoassay based on phage mimotope for nontoxic detection of Zearalenone in maize

Biosynthetic Mycotoxin Conjugate Mimetics-Mediated Green Strategy for Multiplex Mycotoxin Immunochromatographic Assay

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