MICROBIOLOGY Study Guide - Polyclonal Antibodies, Rheumatoid Arthritis, Antiserum

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amigo

4 Jan 2022

School

Kerala University of Health Sciences - KUHS

Department

Pharmacy

Course

MICROBIOLOGY

Professor

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The value of antibodies as tools for locating or identifying antigens is well established. For many years, antiserum extracted from human or animal blood was the main source of antibodies for tests and therapy, but most antiserum is problematic. It contains polyclonal antibodies, meaning it is a mixture of different antigen- specific antibodies, reflecting. The technology for producing monoclonal antibodies involves hybridizing human cancer cells and activated rabbit or mouse b cells in tissue culture systems. Mouse-human chimeric mabs (mouse variable chains on human constant chains) and humanized (mouse hypervariable amino acids in human antibodies) are available. Additionally, human mabs can be made in escherichia coli from bacteriophage containing human. The mabs target various types of cancer, autoimmune diseases, infection, sepsis, rheumatoid arthritis, crohn"s disease, and alzheimer"s disease, to name a few. Additionally, antibody pieces such as (e) the single domain and (f) the single-chain fragments can also be used.

Related Questions

How do you detect a protein by immunoblot (Western blot) using monoclonal antibodies? ELISA assays are often used clinics to detect viral infections. Explain briefly how ELISA assay works.

goldreindeer614

5.. Use your understanding of direct and indirect ELISA to answer the following questions

a. Suppose you work in a clinical lab and are responsible for screening a large array of samples for the presence of gonadotropin (a positive indicator of pregnancy). Which one of the two types of ELISA would be the best approach for this purpose?

b. Suppose you are working with ribonuclease A (RNase A) in the laboratory and that you produce a panel of mouse monoclonal antibodies to RNase A as part of your work. Your next step is to determine which of your monoclonal antibodies is able to recognize RNase A the best. Would you use indirect ELISA or sandwich ELISA for this? You should assume that no other lab has monoclonal antibodies to RNase A (that's why you're trying to make them in the first place!). You may assume that you have significant amounts of RNase A at a known concentration.

Please give good answers with explanations. Thanks so much!!

sapphireporcupine579

I need to make an antibody for the extracellular portion of the Notch receptor. I have made S35 labelled Delta and my boss has given me an antibody that recognizes Delta and is conjugated to a chromophore that gives off light. Place the following statements in order beginning with labeling. HINT: Labeling, protein enrichment, immune response, making monoclonal cell lines, antigen screening of monoclonal cell lines.

A) Hybridoma cells are selected by plating them in HAT buffer.

B) Using direct ELISA you coat individual tissue culture wells/dishes with a single monoclonal antibody.

C) I detect my protein fraction using a geiger counter.

D) You add a solution containing radiolabeled Notch extracellular domain to the wells.

E) You only keep monoclonal hybridoma cell lines that give light. The other cell lines you throw away. You have your antibody!

F) I centrifuge my homogenate in a sugar or salt gradient.

G) I inject a mouse with my protein, giving booster shoots every week.

H) Heterokaryon cells are generated by the fusion of plasma cells and a myeloma cell line.

I) You wash away excess Delta antibody.

J) You add medium containing the Delta antibody that is covalently bound to a chromophore to the individual wells of the ELIZA.

K) To determine if the monoclonal antibody from any given monoclonal cell line recognizes the Notch extracellular domain, you look for light.

L) I use dialysis to remove excess salt and or sugar and to place the protein into a solution that mimics that of the cytoplasm.

M) The mouse is euthanized to harvest the plasma cells in the spleen.

N) Vast quantities of the exported form of the triggered antibody from plasma cells are released.

O) On some B cells, individual antibody receptor forms are bound by the Notch antigen for and activate B cells.

P) I fractionate the proteins.

Q) Individual hybridoma cells are cultured in separate tissue culture wells/dishes to create monoclonal cell lines. These lines export the unique antibody they make into the culture medium.

R) I take culture cells with the Notch receptor expressed on the cell surface and mix them with radio-labelled Delta.

S) B cells divide to give rise to plasma cells and memory cells.

T) You wash away excess Notch extracellular domain from the coated ELISA wells

MICROBIOLOGY Study Guide - Polyclonal Antibodies, Rheumatoid Arthritis, Antiserum

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