ELISAHeri WibowoEnzyme Linked Immuno-Sorbent Assay

DefinisiELISA Test yang dirancang berdasarkan prinsip imunologi (Antigen antibodi) mengunakan label enzim yang dapat ditujukan untuk Mendeteksi keberadaan protein target,Pengikatan enzim / labeling memungkinkan untukMelacak keberadaan protein target (Kwalitatif)Berapa banyak jumlah protein target (Kwantitatif)

Prinsip imunologiAntigenEpitopAntibody

Komponen ELISAPlate mikrotiter /Solid phase portion of assay.Konjugate (suatu ab yang telah berlabel enzim)Subtrate (Substansi yang akan mengatifkan enzim setelah berikatan dg bagian aktif enzim perubahan warna)Bloking Bufer pencuci/deterjen (Washing buffer)

Metode ELISADapat didesain berdasarkan pengikat targetnya :Antigen CaptureAntibody CaptureProsedur deteksinyaDirect ELISA : Bila konjugate (Berlabel enzim) berikatan langsung dengan protein target / antigenIndirect ELISA : Bila konjugate (Berlabel enzim) tidak berikatan langsung dengan protein target / antigen

Protokol Umum ELISADeteksi AntibodiCoatingBlockingConjugateSubtrate

Direct Antigen Capture

Direct antibody capture

Indirect Triple Antibody SandwichIndirect antibody capture

Indirect Antigen Capture

COMPLEMENT FIXATION

Complement FixationPrinciple

In the complement fixation reactin an antigen-antibody reaction is demonstrated by binding of complement.

The reaction can be visualized with the aid of an indicator reactin (hemolytic system).

The hemolytic system consist of sheep erythrocytes and antiserum to sheep erythrocytes (amboceptor).

If no antigen-antibody reaction develops with the consumption of complement, the erythrocyte will be lysis completely and the hemoglobin will be released.

*Cut YasminKonsep Dasar Imunologi