INVESTIGADORES
URRUTIA Mariela
congresos y reuniones científicas
Título:
Generación de anticuerpos inhibitorios de enzimas. Anticuerpos de dominio único inhiben la actividad transialidasa del Tripanosoma Cruzi
Autor/es:
URRUTIA M.
Lugar:
Capital Federal
Reunión:
Exposición; Seminarios del Centro de Estudios Biomédicos, Biotecnológicos, Ambientales y Diagnóstico (CEBBAD).; 2012
Institución organizadora:
Universidad Maimónides/CEBBAD,
Resumen:
SINGLE-DOMAIN LLAMA ANTIBODIES INHIBIT
TRANS-SIALIDASE ACTIVITY OF TRYPANOSOMA CRUZI.
Urrutia M.
Fundación Instituto Leloir. Bs As. Instituto de Investigaciones
Biotecnológicas (IIB), UNSAM. Bs As.
e-mail: murrutia@leloir.org.ar
Trypanosoma cruzi
trans-sialidase (TcTS) constitutes a key
molecule in both the establishment of the infection and the development of
pathologic abnormalities related with Chagas disease.
Llamas
produce, additionally to conventional antibodies, unusual immunoglobulins
devoid of light chains. Their binding site is formed solely by one variable
region (VHH). The binding strategies of
these VHH are very particular, their CDR3 region form long extensions that can
protrude into cavities on antigens, e.g. the active site crevice of enzymes
being suitable for the development of enzyme inhibitors.
The
cDNA isolated from lymphocytes of llamas immunized with TcTS was used to
generate by PCR a VHH library composed of 1x106 clones. Phage
display was used for panning and screening of the library. Preliminary TcTS
inhibition screening allowed us to identify four strong inhibitors clones (SI), which have the same CDR3Â’s length sharing 16 out
of 17 CDRs residues, and few differences along the rest of the sequences.
Inhibitory capacity, affinity measurements and native/denature recombinant
enzyme recognition assays are shown.
We
present different evidences suggesting that the SI could be recognizing an
epitope probably overlapped to the active site of the enzyme.