Antibody Library Construction and Screening Quiz

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What is one key advantage of shuffling the L and H chains in a combinatorial library to obtain Fv molecules?

Lower efficiency in screening bacteriophage plaques

Reduced need for random mutagenesis

Decreased variation in the molecules

Increased recognition of unusual epitopes (correct)

How was the bacteriophage λ vector engineered to address the drawback of lysing bacterial host cells?

Inserting H and L chain DNA sequences into a site flanked by plasmid DNA sequences (correct)

Removing the H and L chain DNA sequences from the vector entirely

Enhancing the protein production capacity of the vector

Increasing the lytic activity of the bacteriophage λ vector

What is a significant benefit of using the phage display technique for antibody production?

Increased need for hybridoma development

Limited variations in gene copy number

No need for animal immunization (correct)

Decreased quantification capabilities

How does the use of plasmids in E.coli enhance antibody production?

Facilitates the production of large numbers of Fv fragments Signup and view all the answers

Why are bacteriophage λ vectors not ideal for large-scale protein production?

They lyse bacterial host cells Signup and view all the answers

In what way does random mutagenesis contribute to achieving even greater variation in combinatorial libraries?

By introducing diverse mutations into DNA sequences Signup and view all the answers

What is the purpose of using two different restriction enzymes in directional cloning?

To create non-complementary sticky ends Signup and view all the answers

How do bacteriophage vectors differ from plasmids in the context of antibody production?

Bacteriophage vectors contain plasmid sequences Signup and view all the answers

What is the number of different antibodies that can be expected from a phage library compared to the mammalian antibody repertoire?

$10^8$ Signup and view all the answers

Why is biopanning used in the context of phage display technology?

To selectively enrich phages that bind to antigens Signup and view all the answers

What is the primary advantage of combining L and H chain cDNAs on one vector in antibody production?

To create a diverse array of unique antibody genes Signup and view all the answers

How does using different restriction enzymes affect the generation of combinatorial antibody libraries?

It creates a random set of libraries for screening Signup and view all the answers

What is the advantage of cloning and expressing complete antibody molecules in E.coli for therapeutic purposes?

Direct screening of E.coli libraries for binding to various antigens Signup and view all the answers

What is the role of the fusion protein that becomes anchored within the inner membrane of E.coli during the expression of antibody molecules?

Facilitates secretion of antibodies into the periplasm Signup and view all the answers

How are full-length antibodies engineered in E.coli libraries to ensure their secretion into the periplasm?

By anchoring them to the inner membrane using a fusion protein Signup and view all the answers

What is the purpose of engineering E.coli to express a fusion protein containing a portion of Staphylococcus aureus protein A?

To facilitate binding of antibodies to antigens Signup and view all the answers

Why is it advantageous for therapeutic antibodies to be full length in many applications?

Increased binding affinity to antigens Signup and view all the answers

What is the key advantage of screening an E.coli library directly for binding to antigens?

Facilitates the selection of antibodies with specified affinities Signup and view all the answers