A Monoclonal Antibody Against F1-F0 ATP Synthase Beta Subunit

Yuan, Jun; Zhang, Cheng; Sui, Fang; Zhuang, Zhenhong; Ling, Sumei; Wang, Shihua

doi:10.1089/hyb.2012.0033

Cited by 9 publications

(7 citation statements)

References 23 publications

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“…Determination of the mAb isotype was identified according to the instruction of mouse Monoclonal Antibody Isotyping (IgA, IgM, IgG 1 , IgG 2a , IgG 2b , IgG 3 ) kit [ 12 ]. Chromosome analysis was identified according to the publication in our lab [ 13 ].…”

Section: Methodsmentioning

confidence: 99%

Effective preparation of a monoclonal antibody against human chromogranin A for immunohistochemical diagnosis

et al. 2018

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BackgroundHuman chromogranin A (CgA) is a ~ 49 kDa secreted protein mainly from neuroendocrine cells and endocrine cells. The CgA values in the diagnosis of tumor, and in the potential role in prognostic and predictive tumor as a biomarker.ResultsThe synthesized gene of CgA coding area was cloned and expressed as fusion protein CgA-His in procaryotic system. Then the purified CgA-His protein was mixed with QuickAntibody-Mouse5W adjuvant, and injected into mice. The CgA-His protein was also used as coating antigen to determine the antiserum titer. By screening, a stable cell line named 4E5, which can generate anti-CgA monoclonal antibody (mAb), was obtained. The isotype of 4E5 mAb was IgG2b, and the chromosome number was 102 ± 4. Anti-CgA mAb was purified from ascites fluid, and the affinity constant reached 9.23 × 109 L/mol. Furthermore, the specificity of the mAb was determined with ELISA, western blot and immunohistochemistry. Results indicated that the mAb 4E5 was able to detect chromogranin A specifically and sensitively.ConclusionsA sensitive and reliable method was successfully developed for rapid production of anti-CgA mAb for immunohistochemistry diagnosis in this study, and the current study also provides conclusive guidelines for preparation of mAbs and implements in immunohistochemistry diagnosis.

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Section: Methodsmentioning

confidence: 99%

Effective preparation of a monoclonal antibody against human chromogranin A for immunohistochemical diagnosis

et al. 2018

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“…Similarly, the production of mAbs requires multiple phases, long duration, and high cost. Currently, mAbs have been produced against a number of mycotoxins such as fumonisin B1 (Yuan et al, 2012;Ling et al, 2014Ling et al, , 2015b, citreoviridin (Jin et al, 2014), marine toxins (Saeed and Wang, 2016), and other exo-and endo-antigens. Similarly, mAbs against transmembrane enzymes have been produced (Yuan et al, 2012).…”

Section: Monoclonal Antibodymentioning

confidence: 99%

Antibody Engineering for Pursuing a Healthier Future

et al. 2017

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Since the development of antibody-production techniques, a number of immunoglobulins have been developed on a large scale using conventional methods. Hybridoma technology opened a new horizon in the production of antibodies against target antigens of infectious pathogens, malignant diseases including autoimmune disorders, and numerous potent toxins. However, these clinical humanized or chimeric murine antibodies have several limitations and complexities. Therefore, to overcome these difficulties, recent advances in genetic engineering techniques and phage display technique have allowed the production of highly specific recombinant antibodies. These engineered antibodies have been constructed in the hunt for novel therapeutic drugs equipped with enhanced immunoprotective abilities, such as engaging immune effector functions, effective development of fusion proteins, efficient tumor and tissue penetration, and high-affinity antibodies directed against conserved targets. Advanced antibody engineering techniques have extensive applications in the fields of immunology, biotechnology, diagnostics, and therapeutic medicines. However, there is limited knowledge regarding dynamic antibody development approaches. Therefore, this review extends beyond our understanding of conventional polyclonal and monoclonal antibodies. Furthermore, recent advances in antibody engineering techniques together with antibody fragments, display technologies, immunomodulation, and broad applications of antibodies are discussed to enhance innovative antibody production in pursuit of a healthier future for humans.

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“…The isotype of 1E6A4 mAb was determined with mouse monoclonal antibody isotyping (IgA, IgM, IgG 1 , IgG 2a , IgG 2b , IgG 3 ) kit. Chromosome analysis was carried out as described previously [12]. In brief, the hybridoma cells were stained with Giemsa stain solution, and the chromosome number was counted under the fluorescence microscope.…”

Section: Methodsmentioning

confidence: 99%

Generation and characterization of a monoclonal antibody against human BCL6 for immunohistochemical diagnosis

Jia

Zhang

Wang³

et al. 2019

PLoS ONE

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Background Human B-cell lymphoma 6 (BCL6) gene, usually coding protein of 706 amino acids, is closely associated with large B cell lymphoma. Researches showed that protein mutation or change of expression levels usually happened in the mounting non-hodgkin lymphoma (NHL). Thus BCL6 is considered to be involved in germinal center (GC)-derived lymphoma. Results The BCL6 1-350 gene codons were optimized for prokaryotic system. After expression of BCL6 1-350 in E . coli , the BCL6 1-350 protein was purified with Ni column. Then the BCL6 1-350 protein, mixing with QuickAntibody-Mouse5W adjuvant, was injected into Balb/c mice. After immunization and cell fusion, a stable cell line named 1E6A4, which can secrete anti-BCL6 antibody, was obtained. The isotype of 1E6A4 mAb was determined as IgG 2a , and the affinity constant reached 5.12×10 10 L/mol. Furthermore, the specificity of the mAb was determined with ELISA, western blot and immunohistochemistry. Results indicated that the 1E6A4 mAb was able to detect BCL6 specifically and sensitively. Conclusions BCL6 1-350 antigen has been successfully generated with an effective and feasible method, and a highly specific antibody named 1E6A4 against BCL6 has been screened and characterized in this study, which was valuable in clinical diagnosis.

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A Monoclonal Antibody Against F1-F0 ATP Synthase Beta Subunit

Cited by 9 publications

References 23 publications

Effective preparation of a monoclonal antibody against human chromogranin A for immunohistochemical diagnosis

Effective preparation of a monoclonal antibody against human chromogranin A for immunohistochemical diagnosis

Antibody Engineering for Pursuing a Healthier Future

Generation and characterization of a monoclonal antibody against human BCL6 for immunohistochemical diagnosis

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