항체 생성 면역 반응 유도를 통해 생성된 항체의 확인과 ELISA를 이용한 정량적 분석

문서 내 토픽

• 1. B-cell development

  Bone marrow에서는 B cell의 조상 세포인 Progenitor B cell이 있는데, 이 Progenitor B cell로부터 mature B cell이 만들어진다. 이 mature B cell은 Peripheral lymphoid organ으로 이동하는데, antigen을 만나지 않아 activate 되지 않은 상태의 B cell인 naive B cell의 90%는 antigen을 만나지 못해 죽고, 10% 정도만 antigen을 만나 activation된다. Activated B cell은 class switching을 통해 Plasma cell이 되어 항체를 생산하며, 일부 세포들은 memory B cell로 분화되어 남는다. B 세포가 양질의 antibody를 생산해내기 위해서는 T helper cell의 도움이 필요하다.

• 2. TD antigen and TI antigen

  B cell에는 TI antigen receptor와 TD antigen receptor가 있다. TI antigen은 TI antigen receptor에 결합하게 되면, 2개의 signal을 보내 B cell이 T helper cell의 도움 없이도 항체인 IgM을 내보내도록 한다. TD antigen이 TD antigen receptor에 결합하게 되면, 하나의 signal만 보내 바로 항체가 secretion되지 않고, T helper cell의 도움을 받아야한다. T helper cell의 CD40 L과 B cell의 CD40이라는 receptor가 결합하여 T helper cell이 B cell에 signal을 보내면 B cell이 activation된다. 이때 activated B cell은 T helper cell로부터 방출된 cytokine의 종류에 따라 각기 다른 개별형으로 전환된다. 이번 실험에서는 TI antigen으로 LPS를 사용하였으며, cytokine으로는 IL-4을 사용하여 IgG의 secretion을 유도하였다.

• 3. ELISA (Enzyme-Linked Immunosorbent Assay)

  ELISA는 enzyme-conjugated antibody를 이용하여 antigen 또는 antibody 등을 검출하고 질적 또는 정량적으로 분석하는 기술이다. ELISA의 종류로는 Indirect ELISA, Sandwich ELISA, Competitive ELISA 등이 있다. 이번 실험에서는 Sandwich ELISA를 이용하여 detection 하고자 하는 antibody인 IgM과 IgG를 확인하고, 정량적으로 분석하였다. Biotin이 붙어있는 detection Ab을 sample에 넣어준 뒤, biotin과 결합하는 Streptavidin-HRP을 넣어주고, enzyme HRP와 반응하는 OPD를 넣어주어 노란색으로 발색 반응을 일으키는 방식으로 실험을 진행했다.

Easy AI와 토픽 톺아보기

• 1. B-cell development

  B-cell development is a complex and crucial process in the immune system. It involves the differentiation of B-cell progenitors into mature B cells capable of producing antibodies. This process is tightly regulated and involves various stages, including early B-cell development in the bone marrow, where B-cell precursors undergo gene rearrangement and selection, and later stages of B-cell maturation and differentiation in the periphery, where B cells encounter antigens and undergo further selection and differentiation. Understanding the mechanisms underlying B-cell development is essential for understanding the adaptive immune response and for developing therapies targeting B-cell-mediated diseases. Ongoing research in this field continues to shed light on the intricate signaling pathways and transcriptional networks that govern B-cell development and function.

• 2. TD antigen and TI antigen

  The distinction between T-dependent (TD) and T-independent (TI) antigens is a fundamental concept in immunology. TD antigens require the help of T cells for B cells to mount an effective antibody response, while TI antigens can stimulate B-cell activation and antibody production without T-cell help. This difference has important implications for the immune response and the development of vaccines and therapies. TD antigens typically induce a more robust and long-lasting antibody response, as they engage both B cells and T cells, leading to the formation of memory B cells and long-lived plasma cells. In contrast, TI antigens can elicit a more rapid but often less durable antibody response, as they primarily activate B cells directly. Understanding the characteristics and mechanisms of TD and TI antigens is crucial for designing effective vaccines and immunotherapies that can elicit the desired immune response.

• 3. ELISA (Enzyme-Linked Immunosorbent Assay)

  ELISA (Enzyme-Linked Immunosorbent Assay) is a widely used analytical technique in immunology, diagnostics, and biomedical research. It is a powerful tool for the detection and quantification of specific proteins, antibodies, or other biomolecules in a sample. The ELISA method relies on the high specificity and affinity of antibody-antigen interactions, where an enzyme-labeled antibody is used to detect the presence and concentration of a target analyte. The versatility of ELISA has made it an indispensable technique in various applications, such as disease diagnosis, drug development, environmental monitoring, and basic research. The ability to perform ELISA in a high-throughput, sensitive, and quantitative manner has contributed to its widespread adoption in both clinical and research settings. Ongoing advancements in ELISA technology, such as the development of more sensitive detection methods and the integration with automated platforms, continue to expand the capabilities and applications of this essential immunoassay technique.