The Preparation and Identification of a Monoclonal Antibody against Domoic Acid and Establishment of Detection by Indirect Competitive ELISA

Saeed, Abdullah F. U. H.; Ling, Sumei; Yuan, Jun; Wang, Shihua

doi:10.3390/toxins9080250

Cited by 19 publications

(14 citation statements)

References 44 publications

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“…Salmonella enteritidis and Salmonella pullorum are two of the most important Salmonella spp. that threaten the poultry industry, and humans are infected by directly or indirectly eating contaminated water and food, which causes great hazard to human public health security (21,22). In our study, a McAb-based competitive ELISA was established to detect O:9 Salmonella infection in chicken.…”

Section: Discussionmentioning

confidence: 99%

A McAb-Based Direct Competitive ELISA to Detect O:9 Salmonella Infection in Chicken

et al. 2020

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Salmonella enteritidis and Salmonella pullorum belonging to Group O 9 Salmonella are major causative agents of infectious diseases in chicken. O 9 antigen as a part of lipopolysaccharide (LPS) is a predominant detected target for Salmonella infection. To identify the infection, an anti-O 9 monoclonal antibody (McAb)-based direct competitive enzyme-linked assay (O 9 Dc-ELISA) was developed after constraints were optimized; the establishment and application of O 9 Dc-ELISA, compared to two commercial kits and plate agglutination test (PAT), showed that O 9 Dc-ELISA could screen out more positive samples than the PAT method could and produce the same agreement rates with commercial kits in terms of sensitivity in addition to strong specificity to clinical serum samples.

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Section: Discussionmentioning

confidence: 99%

A McAb-Based Direct Competitive ELISA to Detect O:9 Salmonella Infection in Chicken

et al. 2020

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“…In order to avoid the false-positive results in screening tests, there is a misconception that heterologous conjugates are necessary for screening of hapten-specific antibody-secreting hybridoma. Accordingly, the hepten-OVA conjugates, for instance, should be used to detect the hapten-specific antibodies, and vice versa (Jin et al 2014; Saeed et al 2017; Gao et al 2007; Esteve-Turrillas et al 2015; Esteve-Turrillas et al 2010). However, in the present study, no hybridoma secreting anti-BSA mAb was screened out using the SAL-BSA conjugate for antibody screening, even though the FBS content in the supernatant was reduced from 10% to 1.0%.…”

Section: Discussionmentioning

confidence: 99%

“…Keyhole limpet hemocyanin (KLH) (Pak et al 1995; Ling et al 2014), bovine serum albumin (BSA) (Kim et al 2004; López-Moreno et al 2014) and ovalbumin (OVA) (Green et al 2012; Parra et al 2014) are the three most common carrier proteins employed for hapten conjugation. In many cases, KLH (or BSA) conjugate is preferred as imunogen for immunization, while BSA (or OVA) conjugate is used as coating antigen to monitor the raised antibodies (Jin et al 2014; Saeed et al 2017; Gao et al 2007; Esteve-Turrillas et al 2015). Hence, BSA or OVA, is often used as control carrier to verify that the antibodies are specific for the target hapten rather than the carrier itself (e.g.…”

Section: Introductionmentioning

confidence: 99%

Bovine serum albumin as an immunogenic carrier facilitating the development of hapten-specific monoclonal antibodies

Zhao

Jiang²

2020

Preprint

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There is a common misconception that the generation of hapten-specific monoclonal antibodies (mAbs) requires the use of a heterologous conjugate to ensure carrier-specific antibodies not being detected. In this study, salbutamol (SAL) was used as a model hapten to exhibit the benefits of bovine serum albumin (BSA) as a carrier for developing hapten-specific mAbs. SAL-BSA conjugate would serve as both an immune antigen and a screening antigen during the preparation of SAL-specific mAbs. Six hybridomas were identified to secret mAbs specific for free SAL with minor or negligible cross-reactivity with other β-agonists. Meanwhile, none of hybrodomas secreting anti-BSA antibodies were screened out even though the fetal bovine serum (FBS) added to the medium decreased from 10% to 1% (v/v). Based on one of the six mAbs, 3F12, a direct competitive enzyme-linked immunosorbent assay (dcELISA) was developed for meausring SAL. Under the optimized assay, the quantitative working range was from 312.5 to 20,000 pg/mL (R2 = 0.9959), with a limit of detection (LOD) of 142.9 pg/mL. The results showed that BSA is an efficient and suitable protein carrier for facilitating the development of hapten-specific mAbs.

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“…As the simplest of the EU regulated toxin groups in relation to the single congener domoic acid to be reported there have been numerous ELISA kits developed for ASP detection [ 81 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 , 110 , 111 ]. Biosense currently has the only commercial ELISA kit for ASP detection that has been validated in single [ 112 ] and multi-laboratory [ 113 ] studies to AOAC standards.…”

Section: Methods For End-product Testing (Ept)mentioning

confidence: 99%

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

Dillon

Zaczek-Moczydlowska

Edwards

et al. 2021

Sensors

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In the past twenty years marine biotoxin analysis in routine regulatory monitoring has advanced significantly in Europe (EU) and other regions from the use of the mouse bioassay (MBA) towards the high-end analytical techniques such as high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS). Previously, acceptance of these advanced methods, in progressing away from the MBA, was hindered by a lack of commercial certified analytical standards for method development and validation. This has now been addressed whereby the availability of a wide range of analytical standards from several companies in the EU, North America and Asia has enhanced the development and validation of methods to the required regulatory standards. However, the cost of the high-end analytical equipment, lengthy procedures and the need for qualified personnel to perform analysis can still be a challenge for routine monitoring laboratories. In developing regions, aquaculture production is increasing and alternative inexpensive Sensitive, Measurable, Accurate and Real-Time (SMART) rapid point-of-site testing (POST) methods suitable for novice end users that can be validated and internationally accepted remain an objective for both regulators and the industry. The range of commercial testing kits on the market for marine toxin analysis remains limited and even more so those meeting the requirements for use in regulatory control. Individual assays include enzyme-linked immunosorbent assays (ELISA) and lateral flow membrane-based immunoassays (LFIA) for EU-regulated toxins, such as okadaic acid (OA) and dinophysistoxins (DTXs), saxitoxin (STX) and its analogues and domoic acid (DA) in the form of three separate tests offering varying costs and benefits for the industry. It can be observed from the literature that not only are developments and improvements ongoing for these assays, but there are also novel assays being developed using upcoming state-of-the-art biosensor technology. This review focuses on both currently available methods and recent advances in innovative methods for marine biotoxin testing and the end-user practicalities that need to be observed. Furthermore, it highlights trends that are influencing assay developments such as multiplexing capabilities and rapid POST, indicating potential detection methods that will shape the future market.

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The Preparation and Identification of a Monoclonal Antibody against Domoic Acid and Establishment of Detection by Indirect Competitive ELISA

Cited by 19 publications

References 44 publications

A McAb-Based Direct Competitive ELISA to Detect O:9 Salmonella Infection in Chicken

A McAb-Based Direct Competitive ELISA to Detect O:9 Salmonella Infection in Chicken

Bovine serum albumin as an immunogenic carrier facilitating the development of hapten-specific monoclonal antibodies

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

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