Monoclonal Antibody Production Methods
48 Questions
0 Views

Choose a study mode

Play Quiz
Study Flashcards
Spaced Repetition
Chat to lesson

Podcast

Play an AI-generated podcast conversation about this lesson

Questions and Answers

What was the main purpose of Koeler and Milstein's experiment?

  • To replace neoplastic cells with B cells
  • To develop a culture medium for T cells
  • To isolate B cells for antibody production
  • To create a stable antibody production system (correct)

    • Why are B cells difficult to maintain in culture?

  • They tend to proliferate uncontrollably
  • They require a specific antigen for activation
  • They lack the necessary nutrients in HAT medium
  • They die without specific activation and survival signals (correct)

    • What type of cells were used alongside B cells to facilitate the experiment?

  • Immortalized B cells
  • Neoplastic plasma cells (correct)
  • Normal T cells
  • Stem cells

    • What does HAT medium provide for the cells in the experiment?

    <p>Survival conditions for fused cells only</p> Signup and view all the answers

    What happens when two B cells are fused?

    <p>The resulting cells die due to lack of survival signals</p> Signup and view all the answers

    What characteristic of cancer cells was critical for the success of the experiment?

    <p>Their immortalization allowing constant proliferation</p> Signup and view all the answers

    What was the role of the specific antigen (protein X) in Koeler and Milstein's method?

    <p>It stimulated antibody production in the B cells</p> Signup and view all the answers

    Which group of scientists was awarded the Nobel Prize for discovering the method used to generate monoclonal antibodies?

    <p>G. Koeler and C. Milstein</p> Signup and view all the answers

    Which suffix characterizes fully human monoclonal antibodies?

    <p>-umab</p> Signup and view all the answers

    What is the primary purpose of phage display in monoclonal antibody production?

    <p>To produce fully human monoclonal antibodies without immunization</p> Signup and view all the answers

    In the phage display process, what is isolated from B cells?

    <p>Genes encoding antibody variable regions</p> Signup and view all the answers

    Which type of monoclonal antibody is characterized by the suffix -ximab?

    <p>Chimeric monoclonal antibodies</p> Signup and view all the answers

    How are the phages that express the desired variable region isolated during phage display?

    <p>Allowing them to kill a bacteria colony expressing a specific antigen</p> Signup and view all the answers

    What is the role of myeloma cells in the production of monoclonal antibodies?

    <p>To immortalize the fused genes for sustained antibody production</p> Signup and view all the answers

    Which of the following is NOT a type of monoclonal antibody indicated in the content?

    <p>Hybridized</p> Signup and view all the answers

    What is the characteristic feature of murine monoclonal antibodies?

    <p>They are designated with the suffix -omab</p> Signup and view all the answers

    What is the primary location of Central Memory T cells?

    <p>In lymphoid organs such as lymph nodes and spleen</p> Signup and view all the answers

    Which sub-population of memory T cells is primarily responsible for rapid initial responses to pathogens?

    <p>Effector Memory T cells</p> Signup and view all the answers

    What effect does falling below the established threshold of memory T cells have on the immune response?

    <p>It prevents a protective reaction from being mounted</p> Signup and view all the answers

    What was the historical significance of Thucydides' observation in 430 BC?

    <p>It suggested the idea of immune memory</p> Signup and view all the answers

    In the context of the immune memory established through observation in the Faroe Islands, what role did age play in susceptibility to measles?

    <p>Older individuals had experienced earlier infections, providing immunity.</p> Signup and view all the answers

    How did immunologists establish the minimum frequency of memory T cells necessary for a protective reaction?

    <p>By counting T cells before, during, and after infection</p> Signup and view all the answers

    What happens to memory T cells after the healing process?

    <p>They persist for future responses.</p> Signup and view all the answers

    Which of the following is NOT a characteristic of Effector Memory T cells?

    <p>Presence mainly in lymphoid organs</p> Signup and view all the answers

    What is the primary purpose of using monoclonal antibodies against CD4 in immunology?

    <p>To isolate and purify specific CD4 T cells</p> Signup and view all the answers

    What technique can be used as an alternative to the depletion of CD4 T cells?

    <p>Transgenic mice lacking CD4 T cells</p> Signup and view all the answers

    What does the 'w' notation signify in the designation CDw186?

    <p>It requires confirmation with a second monoclonal antibody</p> Signup and view all the answers

    How is the expression of a CD molecule represented when a population lacks it?

    <p>CD-</p> Signup and view all the answers

    What is the significance of the terms 'CD80low' and 'CD80high' when referring to dendritic cells?

    <p>They describe the activation status of the cells</p> Signup and view all the answers

    What was the primary outcome of the first International Workshop on Human Leukocyte Differentiation Antigens held in Paris in 1982?

    <p>Deciding the criteria for the CD designation</p> Signup and view all the answers

    Which of the following statements about CD identification is incorrect?

    <p>A unique number is assigned regardless of mAb recognition</p> Signup and view all the answers

    Which method is commonly used for isolating cells besides antibodies?

    <p>Magnetic beads conjugated with antibodies</p> Signup and view all the answers

    What is the primary reason for using mAb in tumor diagnosis?

    <p>To understand the specific subset of cancer</p> Signup and view all the answers

    Which protein is NOT associated with specific subsets of breast cancer?

    <p>p53</p> Signup and view all the answers

    What is the role of mAb in immuno-radiotherapy?

    <p>To bind with radioactive drugs for organ analysis</p> Signup and view all the answers

    Which conjugated form of mAb is preferred for precise targeting in therapy?

    <p>Fab fragment</p> Signup and view all the answers

    What is a significant disadvantage of using Trastuzumab in cancer therapy?

    <p>It is ineffective in cancers that do not express Her2</p> Signup and view all the answers

    How can mAb conjugated with enzymes assist in cancer therapy?

    <p>By catalyzing drug metabolism to reduce resistance</p> Signup and view all the answers

    What characterizes the mAb that are referred to as affinity proteins?

    <p>They include small portions of antibodies used for targeting</p> Signup and view all the answers

    Which form of antibody is described as being composed of three variable regions complexed together?

    <p>Triabody</p> Signup and view all the answers

    What is the primary mechanism through which the Salk vaccine activates the immune response?

    <p>It involves presentation of killed virus peptides via MHCII.</p> Signup and view all the answers

    Which type of immunity is primarily enhanced by the Sabine vaccine's addition of mucosal IgA?

    <p>Mucosal immunity</p> Signup and view all the answers

    Which wild poliovirus strain has been globally eradicated since 1999?

    <p>Type 2</p> Signup and view all the answers

    What is a characteristic of the ideal vaccine discussed in the content?

    <p>Needs to be inexpensive and easily maintained.</p> Signup and view all the answers

    How does the Sabine vaccine contribute to the overall reduction of virus spreading?

    <p>It generates immunity that clears viruses in the intestinal tract.</p> Signup and view all the answers

    Which statement about the mechanism of the Salk vaccine is true?

    <p>It activates T helper cells through MHC II presentation.</p> Signup and view all the answers

    What describes the predominant type of antibody produced first in response to the Salk vaccine?

    <p>IgM</p> Signup and view all the answers

    Which of the following countries had the last reported case of wild poliovirus type 3?

    <p>Nigeria</p> Signup and view all the answers

    Study Notes

    Humoral Response

    • Antibodies are involved in direct and indirect activities
    • Direct activities occur due to antibody binding to antigens in the variable fragments (Fab regions)
      • Toxin neutralization: Antibodies directly bind toxins, inhibiting their activity. This was first demonstrated by Von Behring and Kitasato.
      • Virus and microbe inhibition: Antibodies block the virus from binding to host cells, preventing infection.
      • Immune complex formation: antigen-antibody complexes.
      • Agglutination: Antibody-mediated clumping of targets, often used in blood typing.
      • Receptor and membrane molecule aggregation and internalization: Antibodies bind receptors or membrane molecules, preventing signals from being sent or received.
    • Indirect activities are mediated by the constant fragment (Fc domain) of immunoglobulins
      • Fc receptors (FcR) are used to mediate various biological functions.
        • High-affinity receptors (e.g., CD64): involved in activation, have ITAM sequences
        • Low-affinity receptors (e.g., CD32): may have ITAM or ITIM sequences, some not transducing signals due to lack of ITAM.
        • pH-dependent receptors (e.g., FcRn): transport IgG across the placenta and maintain IgG levels.
    • Immune phagocytosis (opsonization): Antibodies coat microbes, increasing their recognition and uptake by macrophages.
    • Cytotoxic activity by NK cells and other cells: Antibodies binding to an unwanted cell surface lead to cell lysis by NK cells, macrophages, and other cells.
    • Induction of mast cell/granulocyte degranulation: Antibodies trigger release of inflammatory mediators.
    • Complement activation: Antibodies activate the complement system, leading to inflammation and cell lysis via membrane attack complex (MAC)
    • Antibody-dependent cellular cytotoxicity (ADCC): Antibodies bind to infected cells, triggering killing by natural killer cells
    • Antibody feedback: The amount of IgG influences antibody production. FcRIIB binding of immune complexes regulates antibody production.

    Monoclonal Antibodies

    • Monoclonal antibodies (mAbs) are monospecific (recognizing a single epitope)
    • Isolated from a single lymphocyte clone, ensuring consistent action.
    • Their consistent characteristics make them useful for diagnostics and therapy
    • Immunogenicity is an issue if the antibody is from a different species (e.g., murine)
      • This is why research has focused on techniques like production of chimeric or humanized mAbs
    • Various techniques are used to generate monoclonal antibodies
      • Fusion of antibody-producing B cells with immortalized myeloma cells, creating hybridoma cells
      • Using phage display to select antibodies with the desired characteristics
    • The development of mAbs has significantly advanced several biological fields due to
      • Their specific and consistent character
      • Their relatively easy production compared to polyclonal antibodies

    Tumor Diagnosis and Therapy using mAbs

    • mAbs are helpful in tumor diagnosis and therapy
    • mAbs bind to tumor-specific antigens, triggering immune system responses or cell death
      • Increased immunogenicity: mAbs bind to tumor cells, triggering NK cell or macrophage activation
      • Intracellular pathways: mAbs bind to tumor receptors or oncogenes, blocking signals necessary for tumor cell survival or blocking the process
    • Trastuzumab: example of mAb used against Her2 receptor-expressing cancer cells, which inhibits the expression.
    • Several types of mAb therapies face challenges because of differences in structure and half-life when used in humans.
      • Some efforts are made to humanize the antibody fragments (e.g.,CDRs) to reduce the immune response for therapy

    How to Solve Issues with Murine mAbs

    • Chimeric mAbs: Human constant regions are fused to murine variable regions to reduce immunogenicity and improve stability.
    • Humanized mAbs: Only the antigen-binding regions (CDRs) from murine mAbs are retained, with human framework regions, to largely decrease immunogenicity.
    • Fully Human mAbs
    • Other approaches, such as the generation of human mAbs via phage display or transgenic mice, have been used to overcome immunogenicity

    Applications of Monoclonal Antibodies

    • Serological Immunodiagnosis: Used to identify blood types (e.g., ABO) and disease-associated antigens (e.g., HLA).
    • Cancer Diagnosis and Treatment: mAbs are used to detect certain cancer types and act as therapies to engage the immune system to treat conditions and target specific cells.
    • Immuno-radiotherapy: mAbs conjugated to radioisotopes, used in diagnostics to image specific tissues or organs.

    More on mAbs

    • mAbs can be conjugated or fused with various other substances (toxins, enzymes) in order to have different effects, or target them in a controlled way.
    • The choice of the mAb type to perform a certain experiment will depends on the target that needs to be detected and the specific experimental needs, considering advantages and disadvantages of each type.
    • The mAbs are also referred as affinity proteins, since one of the important applications is to detect the presence of a specific type of antigen.
    • Also, they can be used for immunotherapy.

    Studying That Suits You

    Use AI to generate personalized quizzes and flashcards to suit your learning preferences.

    Quiz Team

    Related Documents

    Humoral Response PDF

    Description

    This quiz explores the key concepts and methodologies involved in the production of monoclonal antibodies, focusing on the pioneering work of Koeler and Milstein. It covers the challenges faced in culturing B cells, the importance of HAT medium, and the role of cancer cells in fusion processes, among other related topics. Test your knowledge about the scientific principles behind these groundbreaking techniques!

    More Like This

    Use Quizgecko on...
    Browser
    Open
    Browser
    Browser