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

• Search for proteins by antibody binding to these specific proteins.

Relationship

Synovial Fluid Autoantigen Inmunoproteomic

• 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.

• Martinez Sanchez, Lina Maria; Molecular biology;

7ed, Medellín: UPB, Faculty of medicina; 2012.