MAB 1 Flashcards
(22 cards)
the FAB region is hyper what?
Hypervariable
Definition of Polyclonal AB
Ab from multiple different clones of B cells heterogenous population of AB produced
What process forms polyclonal AB
Rearrangement of AB genes in B precursors
Why can Serum sickness occur in Pab bioproduction?
Name a problem with Pab
HAHA production
short half life span
Define monoclonal AB
AB from single clone of B cells = homogenous molecules w/ 1 antigen specificity
What does Mab allow the production of?
What 3 things can it then be used for?
High pure AB production w/identical paratopes
Treat cancer / diseases / diagnosis
Name 3 advantages of Mab
targets specific antigen = treat easier
Kills target without collateral damage
Can be purified in large quantities free from contamination
Name 3 disadvantages of Mab
High cost / long time to produce / low affinity
Define Hybridoma
Homogenous population of Ab of predetermined antigen specificity by continous culture of fused cell
What are the two “main ingredients” for Hybridoma
What is used to mix the two together allowing fusion?
Ab secreting B cell + Immortal Myeloma Cell
PEG
How much immunogen is used to insert into mouse?
What is also used with it?
50-100ug
Freunds Adjuvant
What does using an immunogen allow
specificity / enhances immunogenicity
Means mouse is immunizied for spleen B cells to be extracted
HAT in hybridoma stands for what?
What does HT allow?
What does A block?
Hypoxanthine Aminopepterin Thymidine (HAT)
HT = purine synthesis via salvage pathway
A blocks de novo purine synthesis
What does B cell have (hybridoma)
HGPRT ?
de nova blocked by A
has salvage but not immortal
HGPRT +
What does Myeloma cell have (hybridoma)
de nova blocked by A
no salvage
HGPRT -
What does a Hybridoma cell have
De nova blocked by A
Has salvage from B cell + Immortal
When AB mRNA is transfected into Phage DNA, what occurs?
AB expressed on surface
Grown in e.coli
Phage cloned into vectors
What phage is used for HIGH AFFINITY AB production?
Why?
fd+M13 w/minor PIII Capsid
5 copies/phage from 1200 AA b Gal peptides
What phage is used for LOW AFFINITY AB production?
Why?
fd+M13 w/Major PVIII capsid
415 copies / T7 Phage
Name 3 advantages of Phage MAB production
Increased speed/production
Increases MAP repertoire
Cheaper w/better ethics
With Transgenic mice, what is the process done?
mouse IG inactivated + transfected w/human IG
What type of AB is produced w/transgenic mice?
What does this process solve?
high affinity human IG Solves Immunogenic mouse protein residues found in MAB hybridoma
