Yintoni eyenza ii-Antibodies zihlale kwi-Nitrocellulose Membrane yecala lokuThintela ukuVavanya?
Inwebu yeNitrocellulose kwi Ukuhamba kamva Uvavanyo luza emva kwesampulu kunye nepadi yokudibanisa. Inwebu yeNitrocellulose ikwabizwa ngokuba yi-substrate membrane kwaye yenye yezona zinto zibaluleke kakhulu kwaye zibuthathaka kwiziphumo zovavanyo olwenziwe Ukuvavanywa kokuhamba kamva. Kwi-Lateral Flow Assay, kunokubakho izinto ezahlukeneyo ezinokusetyenziswa njenge-acetate yeselulosi, i-polyvinylidene fluoride (PVDF), intlawulo eguqulweyo yenayiloni, kunye ne-polyethersulfone (PES), njl.njl. , kunye unyango lweekhemikhali. Iimbumba zeNitrocellulose azinakho ukuhanjiswa ngokulinganayo kwee-pores ngenxa yoko ixesha lokuhamba kwe-capillary likhethwa ngakumbi. Ngaphambi kokuqhubela phambili ekuqiqiseni kubalulekile ukuba wazi malunga nokuhamba kwe-capillary. Ngokusisiseko, ixesha lokuhamba kwe-capillary lilinganiswa ngemizuzwana okanye iisentimitha kwaye lixesha lilonke elithathiweyo ukuba lizaliswe ngokupheleleyo kumtya wovavanyo okanye kwisiXhobo soVavanyo soVavanyo lweLateral.
Inwebu yeNitrocellulose isebenza njengomzimba ophethe ii-antibodies kwisixhobo se-assay sokuhamba komva. Zikwasetyenziselwa injongo yovavanyo lweenkqubo ezahlukeneyo kunye neentlobo zeentsholongwane okt monoclonals kwaye oku kwenzeka njengesiphumo sokuguqulwa kwe-enzymoimmounoassay.
Kwi inwebu ye-nitrocellulose, ii-antibodies zihlala kwaye zidibanise nezinto zebhayoloji nokuba yeyelatex okanye igolide. Iintsholongwane azivumelekanga ukuba zihambe kwaye azinamandla kwi-membrane ye-nitrocellulose ngenxa yezizathu ezahlukeneyo. Njengokuba kukho imigca yovavanyo kunye nemigca yolawulo kwi-assay ye-lateral flow, ii-antibodies zibambekile phakathi kwale migca mibini. Ukujonga umgca wovavanyo, isetyenziselwa ukubopha isampulu yeprotheni ngelixa umgca wokulawula unee-antibodies ezikhethekileyo zohlobo oluthile ezisetyenziselwa ukufumanisa ngakumbi ii-antibodies kwisampulu. Esinye isizathu sokuba ii-antibodies zihlala kwi-membrane ye-nitrocellulose bubungakanani bepore kwifilimu exhasayo. Ifilimu engeyiyo eyomileyo yenwebu ye-nitrocellulose ayivumeli izilwa-buhlungu ukuba zityibilike kunokuba ziphathwe ngokuqinileyo kwaye ke zihlala kwi-membrane. Oku kuyanceda ukunciphisa ukwahluka.
Ezinye izinto ezibalulekileyo ezenza ii-antibodies zihlale kwi-membrane ye-nitrocellulose kukukhethwa kwe-antibody diluent, indlela yokomisa, ukugxila kwe-antibody yokubamba, kunye nesantya sokukhupha i-reagent. Ukomisa ngokufanelekileyo kuyafuneka ukuqinisekisa ukuba ii-antibody zokubamba zifakwe kwi-membrane; nangona kunjalo, ii-membrane ezingasemva zithatha ixesha elininzi ukoma xa kuthelekiswa nee-membrane ezingasekelwanga. Ngaphandle kobunzima, iinkxalabo zomsebenzisi, ukulungiswa kwesampulu, ukugcinwa, ukuphinda kuvele, kunye nokutolikwa kweziphumo okulula ukukusebenzisa, kufuneka kuqinisekiswe ukuba ukuchongwa okunobuthathaka kweeparameter ezichaziweyo kuyaphawulwa. Eminye yale migangatho, efana nokutolikwa kweziphumo kunye novakalelo, ibangelwa kakhulu kukungashukumi kweearhente zokufumanisa kunye nokugcinwa kwendawo kunye neeparameters zendawo yovavanyo.
Enye indlela eluncedo ekwenzeni ii-antibodies zihlale kwi-membrane ye-nitrocellulose kukuncamathelana kwe-antibodies kwi-membrane yeLateral Flow Assay. Kwakhona ukunganyibiliki okungafunekiyo kweentsholongwane ezikwi-membrane ye-nitrocellulose ye-LFA zibonisa amandla amakhulu eembumba ze-nitrocellulose ekubambeni ii-antibodies kuyo. Njengoko sele kukhankanyiwe ngaphambili kwi-Lateral Flow Assay kukho amasuntswana anomdla kwaye abalulekile kuvavanyo lwesampulu. Ukuqhagamshela oku kumasuntswana egolide e-colloidal, zingqinekile ukuba zenze iantigen zihlale kwi-cellulose membrane. Kule nkqubo, inyathelo lokuqala kukudlula kwintengiso yesibulala-mntu efunekayo kumphezulu wenanoparticle yegolide colloidal. Lo ngomnye wemiceli mngeni kuVavanyo lokuHamba kweXesha eliseMva. Ukulungiselela le njongo, amasuntswana okudibanisa ayenziwe ngokulula enza ukuba ii-antibodies, iimolekyuli kunye nezinye izinto zihlale kwi-nitrocellulose membrane.
Oku kukwabonelela ngokuncamathisela okulula kunye nokugqunywa kwamachiza omzimba kumphezulu kubandakanya ezinye ii-analytic kunye ne-biomolecule. Ngaphandle kokuba amaso e-latex kwi-lateral flow assay ayanceda ekwenzeni ii-antibodies zihlala kwi-membrane ye-nitrocellulose. Ukudityaniswa nje kokuhamba kunokunceda ekwenzeni amajoni omzimba ahlala kwi-nitrocelluse membrane yovavanyo lwecala lokuvavanywa kokuhamba. Nangona ezi nkqubo kunye noonobangela zinokwenza ukuba ii-antibodies zihlale kwi-membrane ye-nitrocellulose ye-lateral Flow Assay, zithatha ixesha kunye neenkqubo ezinde kakhulu Ayisiyiyo le kuphela, kodwa ifuna izixhobo ezinkulu kunye nezinye izinto ezizenza zixakeke kwaye ngexabiso. Kwakhona, ezinye izizathu zifuna ukwenziwa kwe-pH esebenzayo kwaye le yinto ebanzi, ke, iindlela ezintsha kunye neenkqubo ziye zafakwa kwifowuni yeLateral Flow Assay ekhokelela ekusetyenzisweni kwezinye iindlela. Isibini esinezixhobo zokulwa umzimba ngokunganyanzelekanga kwi-nitrocellulose ngokufakwa emzimbeni. Ngengcambu ezinzulu kuphando, izilwa-buhlungu zinokuhlala kwi-membrane ye-nitrocellulose ngenxa yokunxibelelana kwe-hydrophobic kunye ne-dipolar phakathi kwemimandla yeeprotheyini kunye ne-nitrocellulose, ekhokelela kulondolozo lwee-antibodies.
isiphelo
Ukuqukumbela nokushwankathela ngokukhankanya le nkcazo ingentla, i-nitrocellulose yinxalenye ebalulekileyo ye-assay flow lateral kunye nesixhobo esisetyenziselwa ukuvavanya. Enye yezona zinto zibalulekileyo nezibalulekileyo ezimisela ukuVavanywa kokuPhuma kweXesha kunye novakalelo lwayo yi-Substrate Membrane okanye Inwebu yeNitcocellulose. Imigca yovavanyo kunye nolawulo ityhilwe ngaphezulu kwenwebu ukuqinisekisa ukubopha okuhle kunye nokubanjwa kwee-antibodies. I-adsorption engachazwanga kuvavanyo nakwimigca yolawulo inokuchaphazela ubuntununtunu beziphumo, zikhokelela kumaxabiso eziphumo ezingezizo. Njengoko i-nitrocellulose yenye yeendawo zokuvavanywa kokuhamba kwelicala apho isampuli idlulisiweyo. Ukusuka kwisampulu, izilwa-buhlungu kunye namalungu e-antigenic kuzo zombini iisampulu kunye nezixhobo ziyahlangana kwaye zidlulisele kwinxalenye elandelayo. Ukusuka apho, isampulu ihamba kwi-membrane ye-nitrocellulose ethi emva koko ithathe ii-antibodies kunye neeantigen ezifunekayo kuvavanyo. Iipropathi kunye nezizathu ezenza ukuhlala kwe-antigen kwi-membrane ye-nitrocellulose kuchaziwe kwinkcukacha engentla kugubungela umxholo wephepha.
Ucaphulo
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