Humanisation of monoclonal antibodies

Question 1

monoclonal antibodies made in mice and bacteria ave to be

Answer 1

disguised as human

Question 2

why do monoclonal antibodies have to be disguised as human

Answer 2

otherwise immune system will recognise them as foreign and destroy them

Question 3

if you put a mouse antibody into a human

Answer 3

around 5 days after it will be destroyed and excited from the system

Question 4

two main reasons monoclonal antibodies have to be humanised

Answer 4

1) immunogenicity

2) Origin

Question 5

immunogenicity issues

Answer 5

most mAbs used in vivo are of mice origin and therefore Xenogenic

Question 6

due to mAbs being xenogenic (of mouse origin)

Answer 6

human anti-mouse antibody (HAMA) reaction occurs

Question 7

HAMA

Answer 7

human antibody recognised the mouse antibody as foreign and destroys it

Question 8

origin of mAbs issue

Answer 8

mouse origin diminishes mAbs ability to elicit effector mechanisms such as complement-mediated lysis or antibody dependent cell-mediated cytotoxicity

Question 9

if antibody is not humanised

Answer 9

will not elicit complement mediated lysis or antibody dependent cell-mediated cytotoxicity

Question 10

use of xenoantibodies in human has proven

Answer 10

severely limiting due to them being seen as foreign proteins by the immune system

Question 11

name 4 strategies used to increase the human content of the mAbs to prevent rejection

Answer 11

1. production of human hybridomas
2. replacement of the consent regions of mouse mAbs with those of human antibodies
3. replacement of CDRs of human antibody with those of a mouse antibody via CDR grafting
   4) production of fully humanised mAbs in transgenic mice

Question 12

Production of human hybridomas

Answer 12

ethical issues and low efficiency

Question 13

Replacement of the constant regions of mouse mAbs (powerful) with those of human antibodies

Answer 13

chimeric antsy
- leaving important VH/VL parts of the Fab fragment from the the mouse–> however will eventually eb recognised by humans

Question 14

Replacement of complementarity- determining regions (CDRs) of human antibody with those of a mouse antibody via CDR grafting

Answer 14

makes humanised antibody

Question 15

CDR grafting

Answer 15

takes a human antibody dn swaps out the CDRs with the CDrs of a mice so that the hyper variable regions are specific for antigen of interest

Question 16

a set of CDRs constitute a

Answer 16

paratope

Question 17

antibody expression in chinese hamster ovary cells

Answer 17

1) take mouse monoclonal cell line and extract mRNA for variable domains
clone into mammalian expression vector and produces a full length human antibody

2) take that vector and transfect into a Chinese Hamster Ovary cell (CHO)
3) these CHO will grow in vitro and will produce human antibodies with CDRs of the mouse..

Question 18

mAbs with Xi in name

Answer 18

chimeric

Question 19

mAbs with Zu in name

Answer 19

humaniZed

Question 20

mAbs with Mu in name

Answer 20

fully human

Question 21

fully human are

Answer 21

phage display