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Question

Mouse IgM (whole molecule) was injected into rabbit to generate antiserum. Which one of the following mouse antibody components (the same mouse from which IgM molecules were used for immunization) has the possibility to be recognized using the rabbit antiserum in Western blotting?

The correct answer is Both IgG F(ab’) 2 fragment and J chain

When a rabbit is injected with Mouse IgM (whole molecule), its immune system recognizes the Mouse IgM as foreign and generates antibodies against it. These antibodies are contained in the rabbit antiserum. Mouse IgM is a large, pentameric molecule made up of multiple heavy chains ($\mu$ chains), light chains (either $\kappa$ or $\lambda$), and a joining (J) chain which links the monomers together. Therefore, the rabbit antiserum will contain antibodies that bind to various parts (epitopes) of the Mouse IgM molecule, including:

  • Epitopes on the $\mu$ heavy chains (constant and variable regions).
  • Epitopes on the light chains ($\kappa$ or $\lambda$) (constant and variable regions).
  • Epitopes on the J chain.

Now, we need to consider which of the given mouse antibody components share epitopes with the original immunogen (Mouse IgM) and would therefore be recognized by the rabbit anti-Mouse IgM antiserum in Western blotting.

Antibody Components Analysis

Let's analyze each option:

  • IgG F(ab')2 fragment: This fragment is derived from Mouse IgG. It contains the variable regions of both the heavy and light chains, and the first constant domain of the heavy chain (CH1), linked together by disulfide bonds, and also the light chains. Mouse IgG, like Mouse IgM, utilizes either $\kappa$ or $\lambda$ light chains. These light chains have shared constant regions across different immunoglobulin isotypes within the same species (mouse, in this case). Since the rabbit antiserum was raised against whole Mouse IgM, it will contain antibodies against the light chains present in IgM. These anti-light chain antibodies will also recognize the light chains present in the Mouse IgG F(ab')2 fragment.

  • J chain only: The J chain is a small protein covalently linked to polymeric immunoglobulins like IgM and IgA. Since Mouse IgM contains a J chain and the rabbit was immunized with the whole molecule, the rabbit will produce antibodies against the Mouse J chain. If J chain is present as a distinct component or within other structures in the mouse sample being blotted, the rabbit antiserum will recognize it.

  • IgG Fc fragment: This fragment is derived from Mouse IgG and contains the constant region domains (CH2 and CH3) of the IgG heavy chain ($\gamma$ chain). The constant regions of IgG heavy chains ($\gamma$) are distinct from the constant regions of IgM heavy chains ($\mu$). Antibodies raised against the $\mu$ chain constant regions in IgM would generally not cross-react significantly with the $\gamma$ chain constant regions in IgG. Therefore, the rabbit antiserum raised against Mouse IgM is unlikely to strongly recognize the Mouse IgG Fc fragment.

  • Both IgG F(ab')2 fragment and J chain: Based on the analysis above, the rabbit antiserum contains antibodies against both the light chain (present in the IgG F(ab')2 fragment) and the J chain. Therefore, the antiserum has the possibility to recognize both components.

Summary of Recognition

The rabbit antiserum was generated against Mouse IgM. Mouse IgM consists of $\mu$ heavy chains, light chains ($\kappa$ or $\lambda$), and a J chain. The rabbit produces antibodies against all these components.

  • The IgG F(ab')2 fragment contains light chains, which are shared with IgM. Thus, anti-light chain antibodies in the antiserum will recognize it.
  • The J chain is part of IgM. Thus, anti-J chain antibodies in the antiserum will recognize it.
  • The IgG Fc fragment contains IgG-specific heavy chain constant regions, which are different from IgM heavy chain constant regions. Thus, recognition is unlikely or minimal.

Therefore, the rabbit antiserum raised against Mouse IgM has the possibility to recognize both the IgG F(ab')2 fragment (via the light chain) and the J chain.

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Important Questions from Immune System

  1. Consider the following statements -

    1. This type of Immunoglobulin is a type of antibody representing approximately 75% of serum antibodies in humans.

    2. It is present in most abundant quantity in the blood circulation

    Which type of immunoglobulin is being talked about?

  2. Allergy and anaphylaxis are due to -

  3. Cells that release histamine and other vasoactive substances in response to allergens are -

  4. Which is not a part of lymphatic system?

  5. Bacterial infections are generally divided into two broad classes: intracellular and extracellular bacterial infections. Given below are some of the properties which are applicable for bacterial infections.

    A. Humoral immune response is the main protective response against extracellular bacteria.

    B. Innate immunity is not effective against intracellular bacterial pathogens.

    C. Bacterial endotoxins do not induce an innate immune response.

    D. Intracellular bacterial infections generally induce a cell-mediated immune response resulting in secretion of cytokines which activate macrophages.

    Which one of the following combination of statements is correct?

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