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Iyini imisebenzi yekhithi ye-ELISA?
2022-12-23
Ikhithi ye-ELISA isuselwe esigabeni esiqinile se-antigen noma i-antibody kanye nokulebula kwe-enzyme ye-antigen noma i-antibody. I-antigen noma i-antibody eboshelwe ebusweni besithwali esiqinile isawugcina umsebenzi wayo we-immunological, futhi i-enzyme ebizwa ngokuthi i-antigen noma i-antibody igcina kokubili umsebenzi wayo wokuzivikela komzimba kanye nomsebenzi we-enzyme. Ngesikhathi sokunquma, isifanekiso esingaphansi kokuhlolwa (lapho i-antibody noma i-antigen kukalwa) sisabela ne-antigen noma i-antibody ebusweni besiphathi esiqinile. I-antigen-antibody complex eyakhiwe kusithwali esiqinile ihlukaniswa kwezinye izinto ezikuketshezi ngokuwashwa.
Ama-antigen anama-enzyme anelebuli noma amasosha omzimba ayengezwa, nawo abophezela kumthwali oqinile ngokusabela. Ngalesi sikhathi, inani le-enzyme esigabeni esiqinile lilingana nenani lezinto ezisesifanekiso. Ngemuva kokwengeza i-substrate yokusabela kwe-enzyme, i-substrate icutshungulwa yi-enzyme ukuze ibe imikhiqizo enemibala. Inani lomkhiqizo lihlobene ngokuqondile nenani lezinto ezihloliwe ku-specimen, ngakho ukuhlaziywa kwekhwalithi noma ubuningi kungenziwa ngokuya ngokujula kombala.
Ukusebenza okuphezulu kwe-catalytic kwama-enzyme kukhulisa ngokungaqondile imiphumela yokusabela komzimba, okwenza ukuhlolwa kube bucayi kakhulu. I-ELISA ingasetshenziswa ukunquma ama-antigen, kodwa futhi ingasetshenziswa ukunquma amasosha omzimba.
Izimiso eziyisisekelo zekhithi ye-ELISA
Isebenzisa ukusabela okukhethekile kwe-antigen kanye ne-antibody ukuxhuma into ku-enzyme, bese ikhiqiza ukusabela kombala phakathi kwe-enzyme ne-substrate ukuze kunqunywe ubuningi. Into yokulinganisa ingaba i-antibody noma i-antigen.
Kukhona ama-reagents amathathu adingekayo kule ndlela yokunquma:
â I-antigen yesigaba esiqinile noma i-antibody (i-immune adsorbent)
â¡ I-enzyme ebhalwe ukuthi i-antigen noma i-antibody (umaka)
⢠i-substrate yesenzo se-enzyme (i-ejenti yokuthuthukisa umbala)
Esilinganisweni, i-antigen (i-antibody) iboshelwa okokuqala kumthwali oqinile, kodwa isawugcina umsebenzi wayo wokuzivikela komzimba, bese kwengezwa i-conjugate (umaka) we-antibody (antigen) kanye ne-enzyme, esagcina umsebenzi wayo wokuqala wokuzivikela komzimba kanye ne-enzyme. umsebenzi. Lapho i-conjugate isabela ne-antigen (i-antibody) kumthwali oqinile, i-substrate ehambisanayo ye-enzyme iyengezwa. Okusho ukuthi, i-catalytic hydrolysis noma ukusabela kwe-REDOX nombala.
Ama-antigen anama-enzyme anelebuli noma amasosha omzimba ayengezwa, nawo abophezela kumthwali oqinile ngokusabela. Ngalesi sikhathi, inani le-enzyme esigabeni esiqinile lilingana nenani lezinto ezisesifanekiso. Ngemuva kokwengeza i-substrate yokusabela kwe-enzyme, i-substrate icutshungulwa yi-enzyme ukuze ibe imikhiqizo enemibala. Inani lomkhiqizo lihlobene ngokuqondile nenani lezinto ezihloliwe ku-specimen, ngakho ukuhlaziywa kwekhwalithi noma ubuningi kungenziwa ngokuya ngokujula kombala.
Ukusebenza okuphezulu kwe-catalytic kwama-enzyme kukhulisa ngokungaqondile imiphumela yokusabela komzimba, okwenza ukuhlolwa kube bucayi kakhulu. I-ELISA ingasetshenziswa ukunquma ama-antigen, kodwa futhi ingasetshenziswa ukunquma amasosha omzimba.
Izimiso eziyisisekelo zekhithi ye-ELISA
Isebenzisa ukusabela okukhethekile kwe-antigen kanye ne-antibody ukuxhuma into ku-enzyme, bese ikhiqiza ukusabela kombala phakathi kwe-enzyme ne-substrate ukuze kunqunywe ubuningi. Into yokulinganisa ingaba i-antibody noma i-antigen.
Kukhona ama-reagents amathathu adingekayo kule ndlela yokunquma:
â I-antigen yesigaba esiqinile noma i-antibody (i-immune adsorbent)
â¡ I-enzyme ebhalwe ukuthi i-antigen noma i-antibody (umaka)
⢠i-substrate yesenzo se-enzyme (i-ejenti yokuthuthukisa umbala)
Esilinganisweni, i-antigen (i-antibody) iboshelwa okokuqala kumthwali oqinile, kodwa isawugcina umsebenzi wayo wokuzivikela komzimba, bese kwengezwa i-conjugate (umaka) we-antibody (antigen) kanye ne-enzyme, esagcina umsebenzi wayo wokuqala wokuzivikela komzimba kanye ne-enzyme. umsebenzi. Lapho i-conjugate isabela ne-antigen (i-antibody) kumthwali oqinile, i-substrate ehambisanayo ye-enzyme iyengezwa. Okusho ukuthi, i-catalytic hydrolysis noma ukusabela kwe-REDOX nombala.
Umthunzi wombala owukhiqizayo ulingana nenani le-antigen (antibody) okufanele likalwe. Lo mkhiqizo onombala ungabonwa ngeso lenyama, isibonakhulu esibonakalayo, isibonakhulu se-electron, ungaphinda ulinganiswe nge-spectrophotometer (insimbi yelebula ye-enzyme). Indlela ilula, ilula, iyashesha futhi iqondile.
Ngaphambilini:Isingeniso sika-Elisa Plate
