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Sagarika Biswas; Saurabh Sharma;Ashish Saroha;D. S.
Bhakuni; Rajesh Malhotra; Muzna Zahur;Michael Oellerich; Hasi
R. Das; Abdul R. Asif.
-Juan Esteban Quiroz Álvarez
-Jose Alejandro Ramirez Quiros.
III semester.
Faculty of medicine.
UPB
March 11/ 2013
• Chronic inflammatory disease.
• Synovitis.
• Joint destruction
peripheral destruction
pain, stiffness, swelling and loss of mobility.
-Changes in immune system.
• Cell Membrane Proteins.
• Function: cell recognition.
• Genes located on
chromosome 6.
• They (immune cells) attack
foreign antigens or
anomalous.
Viscous fluid (Hyaluronic acid and low protein)
Reduces friction between bones
Lubrication to the joints
Nutrients to the articular cartilage
• 131 personas.
• Entre 25-65 años.
• De ellos, 60 se tomaron muestras de
los pacientes con AR (35 mujeres,
25 hombres), 30
• de osteoartritis (OA) (20 mujeres, 10
varones), 01
• trauma de paciente, y 40 muestras
sanas (20 mujeres, 20 varones) se
recogieron como control negativo.
Separación y análisis
de proteinas.1• 1D (Enfoque isoeléctrico)
2• 2D (SDS)
• Sistema Mini Protean II
Detectar proteinas
especificas en una muestra
determinada.
Luego de la electroforesis
Proteinas se transfieren a un filtro
Con Anticuerpos
Estos reaccionan con la
proteina de interes
1.Puntos de proteinas llevados a cubos de geles de 1mm.
2. Se lavo con agua destilada.
3.Se destiño
4. Se deshidrato
ACN (Acetonitrilo) permitió la extracción de péptidos, se seco y
se almaceno en 0.1% de acido fórmico para su posterior
análisis.
Muestras en SDS
Electrotransferencia de proteinas a la membrana
Incubaron con los 5
anticuerpos primarios
Resultados: Con un valor
significativo.
Presencia de autoanticuerpos
contra GFAP y A1BG.
Cuantificar la cantidad de
autoanticuerpos en los pacientes
con AR.
En este caso es un ensayo indirecto : -Luego del lavado…. Se le añade Ig antiglobulina humana . La cantidad del producto es directamente proporcional a la cantidad de Ac presente en la muestra.
• I really enjoyed the seminar because it had a very clinical
approach, in turn feel that certain paragraphs were really
tangled, but overall I loved it.
• Looking to the future is very productive these autoantigens
discovery because it is a disease that occurs much worldwide.
• With these search of autoantigens science progresses every
day, thanks to early detection of certain diseases and their
best treatment.
• The characterization of these proteins in rheumatoid arthritis
will be very useful for understanding the pathogenesis of the
disease.
• The immunoassay, like western blot, have a bigger importance to know the etiology of varietys disease.
• If we recognize the most common autoantigens in AR, we could prevent early the advance of the patology.
• Western blot have a great especificity in the identification of which antigen rected more whit an antibody the plasm.
• Is necesary found others markers of RA, different to the Rheumatoid factor, because it dont express en 1/3 of patients.