Yintoni eyenza ii-Antibodies zihlale kwi-Nitrocellulose Membrane yecala lokuThintela ukuVavanya?

I-Nitrocellulose Membrane kwiLateral Flow Assay iza emva kwesampulu kunye ne-conjugate pad. I-Nitrocellulose membrane ikwabizwa ngokuba yi-substrate membrane kwaye yenye yezona ndawo zibalulekileyo nezinobuthathaka kwiziphumo zovavanyo olwenziwe kwiLateral Flow Assay . KwiLateral Flow Assay, kunokubakho izinto ezahlukeneyo ezisetyenziswayo ezifana ne-cellulose acetate, i-polyvinylidene fluoride (PVDF), i-charge modified nylon, kunye ne-polyethersulfone (PES), njl. Kwi-Nitrocellulose membrane, kukho ubungakanani beembobo ezahlukeneyo, iimbobo, ubukhulu, kunye nonyango lweekhemikhali . Ii-Nitrocellulose membranes azinawo usasazo olulinganayo lweembobo ngoko ke ixesha lokuhamba kwe-capillary likhethwa ngakumbi. Ngaphambi kokuba uqhubekele phambili kwisizathu kubalulekile ukwazi malunga nokuhamba kwe-capillary. Ngokusisiseko, ixesha lokuhamba kwe-capillary lilinganiswa ngemizuzwana okanye iisentimitha kwaye lixesha elipheleleyo elithathiweyo isampuli ukuze lizaliswe ngokupheleleyo kwi-test strip okanye kwi-Lateral Flow Assay Device.

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- membrane ye-nitrocellulose , ii-antibodies zihlala kwaye zidibana nee-biological particles nokuba yi-latex okanye ii-gold particles. Ii-antibodies azivumelekanga ukuba zihambe kwaye azishukumi kwi-membrane ye-nitrocellulose ngenxa yezizathu ezahlukeneyo. Njengoko kukho imigca yovavanyo kunye nemigca yokulawula kwi-lateral flow assay, ii-antibodies zinamathele phakathi kwezi migca zimbini. Xa ucinga ngomgca wovavanyo, isetyenziselwa ukubopha iproteni yesampulu ngelixa umgca wolawulo une-antibodies ezithile zeentlobo ezisetyenziselwa ukufumanisa ngakumbi ii-antibodies kwisampulu. Esinye isizathu sokuba ii-antibodies zihlale kwi-membrane ye-nitrocellulose bubungakanani be-pore yefilimu engasemva. Ifilimu engasemva engenazimbobo ye-membrane ye-nitrocellulose ayivumeli ii-antibodies ukuba zisheleleke endaweni yoko ziphathwa ngokuqinileyo kwaye ngaloo ndlela zihlala kwi-membrane. Oku kunceda ekunciphiseni ukuguquguquka.

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 oku ngokubhekiselele kwinkcazo engasentla, i-nitrocellulose yinxalenye ebalulekileyo yovavanyo lokuhamba kweseli kunye nesixhobo esisetyenziselwa uvavanyo. Enye yezona zinto zibalulekileyo nezibalulekileyo ezimisela uvavanyo lokuhamba kweseli kunye novakalelo lwayo yiSubstrate Membrane okanye iNitcocellulose Membrane . Imigca yovavanyo kunye nolawulo izotywe phezu kweseli ukuqinisekisa ukubopha kakuhle kunye nokubanjwa kwee-antibodies. Ukutsalwa okungachazwanga kwimigca yovavanyo kunye nolawulo kunokuchaphazela uvakalelo lweziphumo, okubangela amaxabiso eziphumo ezingezizo. Njengoko i-nitrocellulose yenye yeendawo zovavanyo lokuhamba kweseli apho isampuli idluliselwa khona. Ukusuka kwisampuli, ii-antibodies kunye neendawo ze-antigenic zombini isampuli kunye nesixhobo ziyadibana kwaye zidlulele kwicandelo elilandelayo. Ukusuka apho, isampuli igeleza iye kwiseli ye-nitrocellulose ethi ngenxa yoko ithathe ii-antibodies kunye nee-antigens ezifunekayo kuvavanyo. Iimpawu kunye nezizathu ezenza ukuba i-antigen ihlale kwiseli ye-nitrocellulose zichazwe kwiinkcukacha ezingentla ngaloo ndlela zigubungela 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