Yini Eyenza Ama-antibody Ahlale Ku-Nitrocellulose Membrane Lateral Flow Flow Assay?
Ulwelwesi lweNitrocellulose ku Ukugeleza Kwangemuva I-Assay iza ngemuva kwesampula ne-conjugate pad. Ulwelwesi lweNitrocellulose lubizwa nangokuthi i-substrate ulwelwesi futhi lungenye yezingxenye ezibaluleke kakhulu futhi ezizwelayo emiphumeleni yokuhlolwa okwenziwe Ukuhlanjululwa Kwesikhathi Esizayo. Ku-Lateral Flow Assay, kungahle kube nezinto ezahlukahlukene ezisetshenziswayo njenge-cellulose acetate, i-polyvinylidene fluoride (PVDF), ukukhokhiswa kwenayiloni okushintshiwe, ne-polyethersulfone (i-PES), njll. Kulwelwesi lweNitrocellulose, kunosayizi ohlukile wepore, i-porosity, ubukhulu , futhi ukwelashwa kwamakhemikhali. Ulwelwesi lweNitrocellulose alunakho ukusatshalaliswa okulinganayo kwama-pores ngakho-ke isikhathi sokugeleza kwe-capillary sikhethwa kakhulu. Ngaphambi kokuqhubekela phambili ekucabangeni kubalulekile ukwazi ngokugeleza kwe-capillary. Ngokuyisisekelo, isikhathi sokugeleza kwe-capillary sikalwa ngemizuzwana noma ngamasentimitha futhi yisikhathi esiphelele esathathwa isampula ukugcwaliswa ngokuphelele emgqeni wokuhlola noma kuDivayisi ye-Lateral Flow Assay.
Ulwelwesi lweNitrocellulose lusebenza njengomzimba ophethe ama-antibody asecaleni le-lateral flow assay device. Zisetshenziswa futhi ngenhloso yokuhlolwa kwezinqubo ezahlukahlukene nezinhlobo zama-antibodies okungukuthi i-monoclonals futhi lokhu kwenzeka njengomphumela wokuguqulwa kwe-enzymoimmounoassay.
Use ulwelwesi lwe-nitrocellulose, amasosha omzimba ahlala futhi ahlangana nezinhlayiya zebhayoloji noma izinhlayiya ze-latex noma zegolide. Ama-antibody awavunyelwe ukunyakaza futhi awanamandla emzimbeni we-nitrocellulose ngenxa yezizathu ezahlukahlukene. Njengoba kunolayini bokuhlola nolayini bokulawula ku-lateral flow assay, amasosha omzimba abambekile phakathi kwale migqa emibili. Uma kucatshangelwa umugqa wokuhlola, usetshenziselwa ukubopha amaprotheni ayisampula kanti ulayini wokulawula unama-antibodies aqondene nezinhlobo ezithile asetshenziselwa ukuqhubeka nokuthola ama-antibody kusampula. Esinye isizathu sokuthi ama-antibody ahlala kulwelwesi lwe-nitrocellulose ngosayizi wepore wefilimu esekelayo. Ifilimu elisekelayo elingahambisi kahle le-membrane ye-nitrocellulose alivumeli ukuthi amasosha omzimba ashibilike kunalokho aphethwe ngokuqinile futhi ngaleyo ndlela ahlale kulwelwesi. Lokhu futhi kusiza ekwehliseni ukwehluka.
Ezinye izinto ezibalulekile ezenza ama-antibodies ahlale kulwelwesi lwe-nitrocellulose ukukhetha i-antibody diluent, indlela yokomisa, ukuhlushwa kwama-antibodies okubamba, kanye nesilinganiso sokusabalalisa ama-reagent. Ukomisa kahle kuyadingeka ukuqinisekisa ukuthi ama-antibody okubamba ayahlanganiswa kulwelwesi; noma kunjalo, ama-membrane angemuva athatha isikhathi esiningi ukoma uma kuqhathaniswa nama-membrane angasekelwanga. Ngaphandle kobunzima, ukukhathazeka komsebenzisi, ukulungiswa kwesampula, ukugcinwa, ukuphindaphindeka, kanye nokuchazwa kwemiphumela okulula ukuyisebenzisa, kufanele kuqinisekiswe njalo ukuthi ukutholakala okubucayi kwamapharamitha acacisiwe kuyashesha. Ezinye zalezi zindinganiso, njengokuchazwa kwemiphumela kanye nokuzwela, kubangelwa kakhulu ukungasebenzi kahle kwama-ejenti okuthola kanye nokugcinwa kwendawo ngamapharamitha endawo yokuhlola.
Enye indlela ewusizo ekwenzeni ama-antibody ahlale kulwelwesi lwe-nitrocellulose ukunamathiselwa kwama-antibody kulwelwesi lwe-Lateral Flow Assay. Futhi, ukungasebenzi okungahlangani kwama-antibody kulwelwesi lwe-nitrocellulose lwe-LFA kusikisela ngamandla amakhulu kakhulu wezingqimba ze-nitrocellulose ekubambeni amasosha omzimba kuwo. Njengoba kushiwo ngenhla ukuthi kwi-Lateral Flow Assay kunezinhlayiya ezizwelayo futhi ezibalulekile ekuhlolweni kwesampula. Ukuxhumanisa lokhu nezinhlayiya ze-colloidal zegolide, kufakazele ukuthi kwenza ama-antigen ahlale kulwelwesi lwe-cellulose. Kule nqubo, isinyathelo sokuqala ukukhishwa kwe-antibody okungenziwanga okudingekayo ebusweni be-nanoparticle yegolide colloidal. Lokhu kungenye yezinselele kakhulu ku-Lateral Flow Assay. Ngale njongo, izinhlayiya zokuhlangana zenziwe kalula ezenza ama-antibody, ama-molecule nezinye izinto zihlale kulwelwesi lwe-nitrocellulose.
Lokhu kunikeza nokunamathiselwa okulula nokugqokwa kwama-antibodies ebusweni kufaka amanye ama-analytic kanye nama-biomolecule. Ngaphandle kwalapho ubuhlalu be-latex ku-lateral flow assay nabo buyasiza ekwenzeni ama-antibody ahlale kulwelwesi lwe-nitrocellulose. Futhi, ukuhlangana okungahambisani nakho kungasiza ekwenzeni ama-antibody ahlale kulwelwesi lwe-nitrocelluse lovivinyo lokuhlolwa kwe-lateral flow. Noma lezi zinqubo nezimbangela zingenza ama-antibody ahlale kulwelwesi lwe-nitrocellulose ye-Lateral Flow Assay, athatha isikhathi futhi izinqubo ezinde kakhulu Hhayi lokhu kuphela, kepha kudinga imishini emikhulu nezinye izinto futhi ezenza kube matasa futhi kakhulu ebizayo. Futhi, ezinye izimbangela zidinga ukwenziwa okusebenzayo kwe-pH futhi lokhu kuyinto ebanzi, ngakho-ke, izindlela ezintsha nezinqubo zifakiwe kudivayisi yeLateral Flow Assay eholela ekusetshenzisweni kwezinye izindlela. Ama-antibodies ambalwa ahlukaniswe ngokukhethekile ku-nitrocellulose ngokumunca umzimba. Njengoba kunezimpande ezijulile ocwaningweni, ama-antibody nawo angahlala kulwelwesi lwe-nitrocellulose ngenxa yokuxhumana okuthile kwe-hydrophobic ne-dipolar phakathi kwazo zombili izizinda zamaprotheni kanye ne-nitrocellulose, okuholela ekulondolozweni kokuma kwama-antibodies.
Isiphetho
Ukuyiphetha nokuyifingqa ngokwencazelo engenhla, i-nitrocellulose iyingxenye ebalulekile ye-lateral flow assay nedivayisi esetshenziselwa ukuhlolwa. Enye yezinto ezibaluleke kakhulu futhi ezibalulekile ezinquma i-Lateral Flow Assay nokuzwela kwayo yi-Substrate Membrane noma Ulwelwesi lweNitcocellulose. Imigqa yokuhlola nokulawula idwetshwa phezu kolwelwesi ukuqinisekisa ukubopha okuhle nokubanjwa kwamasosha omzimba. Ukukhangiswa okungacacisiwe emigqeni yokuhlola nokulawula kungathinta ukuzwela kwemiphumela, okuholele kumanani wemiphumela yamanga. Njengoba i-nitrocellulose ingenye yezingxenye zesilinganiso sokugeleza kwe-lateral lapho isampula lidluliswa khona. Kusuka kusampula, ama-antibody nezingxenye ze-antigenic zombili isampula nedivayisi ziyahlangana futhi zidlulisele engxenyeni elandelayo. Ukusuka lapho, isampula ligelezela kulwelwesi lwe-nitrocellulose oluthatha amasosha omzimba kanye nama-antigen adingekayo ekuhlolweni. Izakhiwo nezizathu ezenza ukuhlala kwe-antigen kulwelwesi lwe-nitrocellulose kuchazwe emininingwaneni engenhla ngaleyo ndlela kumbozwa isihloko sephepha.
Okubhekwayo
https://www.creative-diagnostics.com/Immunochromatography-guide.htm
https://fnkprddata.blob.core.windows.net/domestic/download/pdf/IBS_A_guide_to_lateral_flow_immunoassays.pdf
https://pubmed.ncbi.nlm.nih.gov/6350463/
https://www.bakerlab.org/wp-content/uploads/2015/12/Holstien_Anal_Bioanal_Chem_2015.pdf
https://www.nature.com/articles/s41598-021-87072-7
https://www.researchgate.net/topic/Nitrocellulose