recombinant antibody Flashcards
what is recombinant antibodies
are mono-specific antibodies produced from the recombinant DNA technique
what is the need for recombinant antibodies
- rescue of specificity
- appropriate production of cell line
- humanization
- design of effector function
- functionalization of fragment evolved from phage display
- antibodies generated from isolated cells
why there is need for recombinant antibodies
- to rescue specificity encoded by unstable hybridoma and cell line
- being able to transform mouse antibody into human antibody through in vivo
- designing antibody for function
what is chimeric antibody
is when V domain of mouse antibody is combined by C domain of human antibody
what is CDR grafting antibody
is antibody produced when CDR of mouse is combined with human V domain framework region and C domain
how does humanization occurs
- by chimerization
-CDR grafting - guided selection
what is HAMA (human anti mouse antibody) reaction
is when patient reacted to mouse antibodies as if they are foreign substances
what is the benefit of humanization
reduce HAMA reaction and immunogenicity
what is guided selection
is antibody produced when they select partly human variant in library is combined with one variant of mouse antibody
what problem can humanization cause
immunogenicity is completely avoided because CDR is not only molecule involved in antigen binding so other substance outside the CDR can affect positioning of CDR
can mouse antibody be transformed into human antibody and how
yes, in vivo
what are we looking for in antibody
-binding to cells and kill them after activation of toxin at antibody binding site
- activate other cells at antibody binding site
- target CD8+ T cells to killed cells
- avidity
-Fc receptor binding
-stability on mucosa/IgA
-absence of effector function
-bi-specific antibody
how does T cells kills cells that are affected by virus
T cells kills cells via antibody specificity rather than by TCR
what is Intact antibodies
is a full antibody
what is a fragment
small part of antibody
what is bi specific antibody fragment
is an antibody that cross link two different epitope
hat is the benefit of bi specific antibody fragment
it has ability to cross link two different cells: CD8+ cytotoxic T cells and bacteria or cancer cell which induce or promote virus cell killing
what is the Systems for production of recombinant antibodies and fragments
- in vitro: no glycosylation
- Escherichia coli : only fragments and no glycosylation
-Pichia pastoris : only fragments
insect cells
myeloma cells
CHO cells
COS cells
HEK293 cells
animals and plant
what are the two important carbohydrate on proteins/antibody
O-linked and N-linked
can carbohydrate in antibodies has important function if they are found on variable domain
yes, the specificity
what does carbohydrate found on antibody affects
- in constant domain it affects effector function
- in variable domains it affects specificity
what is the important aspects of recombinant antibody
is the nature of their carbohydrates
