Kmk

Jhkjjk

gjghjj

Hhhh

Monoclonal antibodies have proven to be attractive reagents which revolutionize how diseases are
diagnosed and treated.

The power of monoclonal antibodies comes from the fact the monoclonal antibodies could bind its
targets with high specificy which allows specific targeting of carbohydrates, peptides, proteins and
polymers.

classically, antibodies are produced in mammalian after immunization with the target of interest and
then hybridoma cells are produced by fusion between specific B cells and myeloma fusion partner. And
then specific hybridoma are selected which produce secreted antibodes specifically against the target of
interest.

A major drawback of this method is the inability to produce antibodies with high affinity against
carbohydrate antigens or conserved antigens.

Carbohydrate antigens are considered as T cells independent antigens because they can’t be presented
by the T cell MHC receptor. B cells secrete antibodies and express on their surfaces antibodies of the
same specificy , when carbohydrate antigens bound on the B cells surface, they internalize and unlike
proteins it can’t be digested an presented on the surface MHC. If peptides are presented on MHC, T cells
bind through TCR and CD40 ligand, this CD40 ligand from T cells allow B cells to express AID enzyme
which mutates the immunoglobulin genes and create a small library with each contains around single
mutation. And then with multiple rounds of selection, only B cells with high affinity can be selected and
amplified.

Globo H is a carbohydrate structure with very important clinical implications, glob H over expression
have been observed in many cancer types including … while in normal conditions it is only expressed in
the epical surfaces of secretory epithelial cells which is not accessible to the blood circulation.

Before we succeeded to change the properties of anti PEG antibodies by expressing AID in hybridoma.

to So far only two hybridomas are produced against globo H and both of them have low affinity, we
thought that if we express AID directly in hybridoma we could mimic the process of somatic
hypermutaions and with repeated rounds of sorting we assumed that we will be able to isolate
improved clones.

Low affinity IgM is a common problem encountered with immunization with carbohydrate antigens.

Another group succeeded to do affinity maturation of antigens by expressing AID in cells that produce
the gene of interest in retrovirus backbone.

Vaccination against carbohydrate antigens by conjugation to a carrier protein seems to be interesting

idea which benefit from the concept of linked recognition, but seems to have no significant beneficial
effect.

The only way which might work is the phage display which enables screening from a huge library but has
the problem of developability