What In the World Is a Monoclonal Antibody?. Antibodies x Antigens. Antibodies . Polyclonal . Monoclonal . Individual B lymphocyte hybridoma is cloned and cultured. Secreted antibodies are collected from culture media. Antibodies that are collected from sera of exposed animal.
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Individual B lymphocyte hybridoma
is cloned and cultured.
Secreted antibodies are collected
from culture media
Antibodies that are collected
from sera of exposed animal
multiple antigenic sites
of injected biochemical.
ONE antigenic site
of injected biochemical
Populações homogêneas de anticorpos derivados de uma única célula produtora de anticorpo, na qual todos os anticorpos possuem a mesma especificidade para um determinante antigênico.
Anticorpos monoclonais -1975
Prêmio Nobel - 1984
A fusion is done when the IgG level is high and the IgM level is low.
Sometimes additional testing is done (Western blots, immunoflurorescence) to determine whether the serum response is specific for the selected antigen.
The mouse with the strongest, most specific response is chosen for the fusion.Test the serum - Decision time
Anticorpos Monoclonais - Produção
2) Media is added to the flasks until the desired volume of antibody-containing supernatant is obtained.
3) When the desired volume is reached, the supernatant is refrigerated and tested by ELISA.
4) If the testing is successful, the supernatant can be used by the investigator, or concentrated and purified.Harvest the antibody
in vitro material is less concentrated
contain high [mAb]
and minor contamination
with mouse Ig
2005: 37.934 articles
can by physically separated
coupled to magnetic beads
Which human cell type
do you want to isolate?
Anticorpos Monoclonais - Utilidades
Common side effect –
Vascular leak syndrome.
Could be relieved
by usage of mutated toxin
Bunch of other toxins
could be used
single chain ribosome inactivating proteinSaponin (from soapwort)
For T-cell leukemia could be
targeted via CD7
Monoclonal antibodies, which are specific for a certain antigen, i.e. prostate carcinoma cells, can be tagged with a radioactive tracer,which when injected into the body will localize in areas of these recurrent carcinoma cells
OctreoScan in a patient
with metastatic small cell carcinoma of the prostate
Anticorpos monoclonais humanos
Falta de linhagens de células de mieloma humano bem estabelecida
Mieloma secretores de Ig
Anticorpos monoclonais humanos
DIFICULDADES (linfócitos B):
Obtenção das células B antígenos-específicas ativadas
sangue x baço/linfonodos
Alternativa: imunizar indivíduos antes de retirar os linf. B
Estimular células B in vitro com Ag
produção de IgM de baixa afinidade
Transferência de linfócitos humanos para SCDI mice
Transformação de linfócitos B com Epstein-Barr
Clone those genes
and express them
as a recombinant protein
in the protein expression system
Sales of Rituxan/
MabThera by Year
Mouse antigen-interacting sequences
Anti-CD20 MAbs (Rituxan) for treatment of non-Hodgkin’s lymphoma
The size and specificity
of the ScFv fragment
may allow for attachment
to cryptic sites
protocols/schema.jpgSelection of antibodies from phage display libraries:
the enrichment of
phage-antibody (circle) on a background of
from serum of cancer patients
XenoMouse genetically engineered
To replace murine IgH and Igk loci
with their human Ig counterparts on YAC
Human Ig transgenes
carry the majority
of the human variable repertoire
and can undergo
from IgM to IgG isotypes.
Donor T cells cause graft vs host (GvH) reaction.
Donor T cells eliminated using anti-CD3 Mabs (OKT3) Muromonab; .
Treat patients with anti-CD3 after solid organ transplants.Unconjugated MAbs as a molecular drugs
2003: 9 AcMo quiméricos ou humanizados aprovados pelo FDA
Leucemia, linfoma não-Hodgkin, rejeição de órgãos, doenças auto-imunes e câncer de mama
RA: artrite reumatóide; MS: esclerose múltipla