Purification of IgA Monoclonal Antibody for Mycobacterium tuberculosis
Journal name: The Malaysian Journal of Medical Sciences
Original article title: Purification of an IgA Monoclonal Antibody Specific for the Acr Protein of Mycobacterium tuberculosis by Immunoaffinity Chromatography
The Malaysian Journal of Medical Sciences (MJMS) is a peer-reviewed, open-access journal published online at least six times a year. It covers all aspects of medical sciences and prioritizes high-quality research.
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Fátima REYES, Oscar OTERO, Frank CAMACHO, María Elena SARMIENTO, Armando ACOSTA
The Malaysian Journal of Medical Sciences:
(A peer-reviewed, open-access journal)
Full text available for: Purification of an IgA Monoclonal Antibody Specific for the Acr Protein of Mycobacterium tuberculosis by Immunoaffinity Chromatography
Year: 2013
Copyright (license): CC BY 4.0
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Summary of article contents:
Introduction
Tuberculosis (TB) remains a significant global health challenge, second only to HIV/AIDS as the leading cause of death from a single infectious agent. Effective control of TB is complicated by the emergence of multidrug-resistant strains and the inconsistent efficacy of the only available vaccine, the Bacillus Calmette-Guérin (BCG) vaccine. Recent research has started to examine the role of antibodies in combating TB, particularly focusing on a monoclonal antibody (mAb) called TBA61, which is specific to the Acr protein of Mycobacterium tuberculosis. Previous studies have demonstrated TBA61's protective qualities in various challenge models, indicating its potential for use in immunotherapy against TB.
Purification of TBA61 mAb using Immunoaffinity Chromatography
A critical advancement in the application of TBA61 mAb is the development of a purification method utilizing immunoaffinity chromatography with the Acr protein as a specific ligand. This method significantly streamlines the purification process compared to traditional approaches, which often require multiple chromatographic techniques to achieve high purity. In this study, the Acr protein was expressed in E. coli and coupled to a cyanogen bromide-activated Sepharose 4B matrix, allowing for the efficient one-step purification of TBA61 from mouse ascites fluid. The results showed an average recovery of 1.46 mg per mL of ascites, and the purified mAb maintained high specificity and reactivity against the Acr protein, confirmed by ELISA and western blot assays.
Conclusion
The introduced immunoaffinity chromatography method for purifying TBA61 mAb is not only rapid and straightforward, but also effective in yielding high-purity antibody in substantial recovery amounts. This streamlined approach serves as a vital advance for future experimental and clinical evaluations of TBA61 mAb's therapeutic potential against TB. The study underscores the potential for leveraging specific antibody-antigen interactions to achieve efficient purification processes, ultimately enhancing the development of targeted immunotherapies for infectious diseases like tuberculosis.
FAQ section (important questions/answers):
What is the purpose of the TBA61 monoclonal antibody?
The TBA61 monoclonal antibody is specific for the Acr protein of Mycobacterium tuberculosis. It has been shown to promote granuloma formation and provides protection against tuberculosis in various animal models.
What methods were used to purify the TBA61 antibody?
To purify the TBA61 antibody, immunoaffinity chromatography was employed using recombinant Acr protein as a ligand. This method allows for rapid and specific purification in a single step, yielding high purity levels.
How effective is the purification method for TBA61 mAb?
The purification method demonstrated an average recovery of 1.46 mg per mL of ascitic fluid. The purified mAb retained its reactivity against the Acr protein, confirming the method's effectiveness for obtaining high yields.
What challenges are associated with purifying IgA monoclonal antibodies?
Purifying IgA monoclonal antibodies traditionally requires multiple methods for satisfactory purity. The similarity in physical characteristics between contaminants and the target can complicate the purification process, necessitating well-chosen chromatographic techniques.
Glossary definitions and references:
Scientific and Ayurvedic Glossary list for “Purification of IgA Monoclonal Antibody for Mycobacterium tuberculosis”. This list explains important keywords that occur in this article and links it to the glossary for a better understanding of that concept in the context of Ayurveda and other topics.
1) Purification:
Purification refers to the process of isolating a specific substance from a mixture, achieving a high degree of purity. In the context of the study, immunoaffinity chromatography was employed for the purification of the TBA61 monoclonal antibody, demonstrating an effective method for obtaining pure antibodies for scientific evaluation and further medical applications.
2) Tuberculosis:
Tuberculosis (TB) is a severe infectious disease caused by the bacterium Mycobacterium tuberculosis. The abstract discusses the implications of TB regarding its treatment challenges, including multidrug resistance and the need for new vaccines, highlighting TB's status as a leading infectious disease globally.
3) Table:
The table presented summarizes the recovery from different cycles of the immunoaffinity purification process, detailing elution volumes, protein content, and calculated recovery rates. This data provides a visual representation of the purification efficiency achieved through the methodology described in the study.
4) Purity:
Purity in the context of this study reflects the quality of the purified TBA61 monoclonal antibody, which was assessed using techniques like SDS-PAGE and Western blot. High purity levels are crucial for reliable scientific experiments and therapeutic applications, ensuring that contaminants do not affect the results.
5) Disease:
The term disease refers to any condition that impairs normal functioning. The discussion covers the disease tuberculosis, emphasizing its global impact and the challenges posed by drug-resistant strains, underscoring the necessity for effective treatments and immunological responses against such diseases.
6) Transmission:
Transmission refers to the spread of an infectious agent. The text highlights the transmission of tuberculosis and the urgency to find effective solutions due to the risks of spreading multidrug-resistant strains, emphasizing the ongoing public health challenge posed by TB transmission.
7) Panda (Pamda):
[see source text or glossary: #Panda#]
8) Study (Studying):
A study is a systematic investigation aimed at discovering or verifying knowledge. This study focused on the purification of the TBA61 monoclonal antibody and its implications for treating tuberculosis, illustrating the scientific inquiry into respiratory diseases and immunological treatments.
9) Drug:
Drugs refer to substances used to treat, cure, or prevent diseases. The context here involves discussing drug treatment for tuberculosis, particularly in relation to multidrug resistance, highlighting the crucial role of effective drug therapies in controlling TB epidemics.
10) Allergy:
[see source text or glossary: #Allergy#]
11) Sharman (Sarma, Sharma, Sarman):
[see source text or glossary: #Sharma#]
12) Nature:
Nature refers to the intrinsic qualities or characteristics of something. In the study, the nature of immunological responses to tuberculosis is explored, examining how specific antibodies interact with the pathogen, reflecting the complex biological interactions in disease processes.
13) Blood:
Blood is the bodily fluid responsible for transporting nutrients, gases, and waste. In the context of the study, the reactivity assays and antibody characterizations emphasize the importance of understanding immune responses in serum or blood to evaluate therapeutic effectiveness.
14) Measurement:
[see source text or glossary: #Measurement#]
15) Transformation (Transform, Transforming):
[see source text or glossary: #Transformed#]
16) Discussion:
Discussion refers to an analytical dialogue about findings or implications. The discussion section in the study interprets the results of antibody purification and its relevance for tuberculosis therapy, examining the implications of the findings for future research and treatment development.
17) Science (Scientific):
Scientific pertains to the systematic study of the structure and behavior of the physical and natural world through observation and experimentation. The purification of TBA61 mAb utilized scientific methods to enhance tuberculosis research, showcasing the necessity of rigorous scientific inquiry in healthcare.
18) Resolving:
[see source text or glossary: #Resolving#]
19) Account:
[see source text or glossary: #Account#]
20) Rules:
[see source text or glossary: #Rules#]
21) Hand:
[see source text or glossary: #Hand#]
22) Milk:
[see source text or glossary: #Milk#]
23) Post:
[see source text or glossary: #Post#]
24) Viru:
[see source text or glossary: #Viru#]
Other Science Concepts:
Discover the significance of concepts within the article: ‘Purification of IgA Monoclonal Antibody for Mycobacterium tuberculosis’. Further sources in the context of Science might help you critically compare this page with similair documents:
Purification method, Polymerase chain reaction, Experimental evaluation, Intranasal administration, Escherichia coli, Positive control, Electrophoresis, Enzyme-linked immunosorbent assay, Protective effect, Negative control, Gel filtration, Structural proteins, Granuloma formation, SDS-PAGE analysis, SDS-PAGE, Protein concentration, BCG vaccine, Mycobacterium tuberculosis, Tuberculosis (TB), Multidrug-resistant Strains, Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis, Monoclonal antibodies, Western blot procedure, Infectious agent, Bioactive Form, Protective Efficacy, Recombinant protein, Monoclonal antibody, Ion exchange, Ascites Fluid, Viral replication inhibition, Immunoaffinity Chromatography, Fusion protein, Acr protein, Acr protein of MTB, Western blot, HspX gene, Recombinant Acr, Anti-IL-4 polyclonal antibody, Therapeutic coverage, Antibody purification, Polysaccharide Capsule, ELISA assay, TB vaccine, Recombinant plasmid, Primary antibody, Combined immunotherapy, Heavy chain-specific anti-mouse IgA, Missouri, USA, 0.1 M citrate buffer, Peroxidase reaction, NCBI BLAST software, Immunopurification of TBA61 mAb, Coupling efficiency, IgA mAb isotypes, Celestial protein, Chromatographic methodologies, TBA61 purification protocol, Co-purification of host immunoglobulins, High purity levels, Immunospecific purification methods, Affinity chromatography handbook, Antigen binding properties, Murine immune response, Monoclonal IgA antibody, Electrophoretic transfer, Bacterial inclusion body proteins, Autoantibodies in anemia, Vaccines conjugate vaccine, Serotyping reagents, IgM and IgA, Nonneutralizing IgA mAb, Analysis of monoclonal antibodies, Antibody purification handbook, Thymus-independent antigen, Thymus-dependent toxoid conjugate, Diagnostic antigens, Secretory immunoglobulin A, IgA isotype, TBA61 mAb, RAcr protein, Purified TBA61, Immunopurification cycles, Proteins from polyacrylamide, Purified immunoglobulin A, Monoclonal antibodies generated, Monoclonal IgA antibodymediated, Rotavirus VP6-specific IgA, IgM and IgA anti-erythrocyte, Monoclonal antibodies of IgA, Specific diagnostic antigens, Anisakis simplex larval antigen, Monoclonal antibodies reactive, Recombinant proteins in Escherichia, Inclusion body proteins using, IgA antibody purification, Transfer of proteins, IgA antibody, Rotavirus replication, Anisakis simplex larval, EBNA1, Inclusion body proteins, Bacterial inclusion body.