Zomwe Zimapangitsa Kuti Ma antibodies Apitirize Kugwiritsa Ntchito Nitrocellulose Kakhungu Kotsatira Kuyenda Kwawo?
Kakhungu ka Nitrocellulose mu Otsatira Otsatira Assay amabwera pambuyo poyeserera ndikuphatikizira pad. Nitrocellulose nembanemba amatchedwanso gawo lapansi nembanemba ndipo ndi chimodzi mwa zinthu zofunika kwambiri ndi tcheru kwa zotsatira za mayeso anachita pa Kufufuza Kwotsatira. Mu Lateral Flow Assay, pakhoza kukhala zida zosiyanasiyana zomwe zingagwiritsidwe ntchito monga mapadi acetate, polyvinylidene fluoride (PVDF), kulipiritsa nayiloni yosinthidwa, ndi polyethersulfone (PES), ndi zina zotero. , ndi mankhwala mankhwala. Nitrocellulose membranes alibe yogawa kofanana ka pores chifukwa chake capillary flow time imakonda kwambiri. Musanapitirire kulingalira ndikofunikira kudziwa za capillary flow. Kwenikweni, nthawi yoyenda ya capillary imayesedwa mumasekondi kapena masentimita ndipo ndi nthawi yonse yomwe chitsanzocho chidatenga kuti chidzazidwe kwathunthu pamayeso oyesera kapena Lateral Flow Assay Device.
Nitrocellulose membrane imagwira ntchito ngati chonyamulira cha ma antibodies omwe amagwiritsidwa ntchito poyeserera. Amagwiritsidwanso ntchito pofuna kuwunika njira zosiyanasiyana ndi mitundu ya ma antibodies ie monoclonals ndipo izi zimachitika chifukwa cha kusinthidwa kwa enzymoimmounoassay.
pa nembanemba wa nitrocellulose, ma antibodies amakhala ndipo amalumikizana ndi tinthu tating'onoting'ono mwina tomwe timakhala ndi tinthu tating'onoting'ono kapena golide. Ma antibodies saloledwa kusuntha ndipo alibe mphamvu pa nembanemba ya nitrocellulose chifukwa cha zifukwa zosiyanasiyana. Popeza pali mizere yoyesera komanso mizere yolamulira panjira yoyeserera, ma antibodies amakhala omangika pakati pa mizere iwiri iyi. Poganizira mzere woyeserera, imagwiritsidwa ntchito popanga puloteni yoyeserera pomwe mzere wolamulira uli ndi ma antibodies omwe amagwiritsidwa ntchito kupezanso ma antibodies omwe ali mchitsanzocho. Chifukwa china chomwe ma antibodies amakhala pamphuno ya nitrocellulose ndikukula kwa pore kwa kanema wothandizira. Kanema wothandizila wosavulaza wa nembanemba ya nitrocellulose salola ma antibodies kuti achoke m'malo mwake amayang'aniridwa mwamphamvu motero amakhala pa nembanemba. Izi zimathandizanso kuchepetsa kusiyanasiyana.
Zinthu zina zofunika zomwe zimapangitsa kuti ma antibodies akhalebe pa nembanemba ya nitrocellulose ndi kusankha kwa antibody diluent, njira yowumitsa, kuchuluka kwa ma antibodies, komanso kuchuluka kwa reagent yomwe imaperekedwa. Kuumitsa bwino ndikofunikira kuti zitsimikizire kuti ma antibodies ogwidwa alumikizidwa ku nembanemba; komabe, nembanemba zakumbuyo zimatenga nthawi yayitali kuti ziume poyerekeza ndi nembanemba zosagwidwa. Kupatula kusinthasintha, nkhawa za ogwiritsa ntchito, kukonzekera zitsanzo, kukonza, kuberekanso, komanso kutanthauzira zotsatira kosavuta kugwiritsa ntchito, nthawi zonse ziyenera kutsimikiziridwa kuti kuzindikira kwa magawo omwe atchulidwa kuyenera kudziwika bwino. Zina mwa miyezo iyi, monga kutanthauzira zotsatira ndi kukhudzidwa, zimakhudzidwa kwambiri ndi kulephera kwa zinthu zozindikira komanso kusunga malo ndi magawo a malo oyesera.
Njira ina yomwe imathandizira kupanga ma antibodies kuti akhalebe pa membrane ya nitrocellulose ndikulumikizana kophatikizana kwa ma antibodies ku nembanemba ya Lateral Flow Assay. Komanso kusakhazikika kwa ma antibodies omwe ali pachimake pa nitrocellulose nembanemba ya LFA kumalimbitsa mphamvu yamankhwala a nitrocellulose pomanga ma antibodies pamenepo. Monga tafotokozera kale kuti mu Lateral Flow Assay pali ma particles omwe ali ofunikira komanso ofunikira pakuyesa kwachitsanzo. Kuphatikiza izi ndi tinthu tamagolide ta colloidal, zatsimikizira kuti ma antigen amakhalabe pakhungu la cellulose. Pochita izi, gawo loyamba ndikumangika kwa antibody wofunikanso pamwamba pa nanoparticle ya golide colloidal. Ichi ndi chimodzi mwazovuta kwambiri pakuyesa kwa Lateral Flay. Pachifukwa ichi, tinthu tating'onoting'ono tomwe timapanga timene timapangitsa kuti ma antibodies, mamolekyulu ndi zinthu zina zikhalebe pakhungu la nitrocellulose.
Izi zimaperekanso kuphatikiza kosavuta ndikuphimba kwa ma antibodies kumtunda kuphatikiza ma analytics ena komanso ma biomolecule. Kupatula kuti mikanda ya latex mumayendedwe ofananira amathandizanso pakupanga ma antibodies kuti akhalebe pa membrane wa nitrocellulose. Kuphatikizanso pang'ono kungathandizenso pakupanga ma antibodies kuti akhalebe pa nitrocelluse membrane of the lateral flow assay test. Ngakhale izi zimayambitsa ndikupangitsa ma antibodies kukhala pa nitrocellulose nembanemba ya Lateral Flow Assay, imatenga nthawi yayitali komanso njira yayitali Osati izi zokha, zimafunikira zida zazikulu ndi zinthu zina zomwe zimapangitsa kuti zikhale zotanganidwa komanso okwera mtengo. Komanso, zina mwazimene zimafunikira kukhathamiritsa kwa pH ndipo ichi ndichinthu chachikulu, chifukwa chake, njira ndi njira zatsopano zaphatikizidwira mu chipangizo cha Lateral Flow Assay chomwe chimapangitsa kugwiritsa ntchito njira zina. Ma antibodies angapo amapangika makamaka pa nitrocellulose chifukwa choyamwa. Ndi mizu yakuya pakufufuza, ma antibodies amathanso kukhala pachimake cha nitrocellulose chifukwa cha kulumikizana kwapadera kwa ma hydrophobic ndi dipolar pakati pamadongosolo onse a protein ndi nitrocellulose, zomwe zimapangitsa kuti chitetezo cha ma antibodies chisungidwe.
Kutsiliza
Kuti mumalize ndikufotokozera mwachidule malinga ndi zomwe tafotokozazi, nitrocellulose ndi gawo lofunikira pakuyesa kotsatizana ndi chida chomwe chimayesedwa. Chimodzi mwazinthu zofunika kwambiri komanso zofunika kudziwa kuti Lateral Flow Assay ndikumverera kwake ndi Substrate Membrane kapena Kakhungu ka Nitcocellulose. Mizere yoyesa ndi kuwongolera imakokedwa pamwamba pa nembanemba kuti zitsimikizire kumangirira ndikulanda ma antibodies. Kutsatsa kwapadera pamayeso ndi mayendedwe olamulira kumatha kukhudza chidwi cha zotsatirazo, zomwe zimabweretsa zotsatira zabodza. Popeza nitrocellulose ndi gawo limodzi mwazomwe zimayendera pambuyo pake. Kuchokera pachitsanzocho, ma antibodies ndi antigenic a zitsanzo ndi zida zake amalumikizana ndikupitilira gawo lotsatira. Kuchokera pamenepo, chitsanzocho chimapita ku nembanemba ya nitrocellulose yomwe imatenga ma antibodies ndi ma antigen omwe amafunika kuti ayesedwe. Katundu ndi zifukwa zomwe zimapangitsa kuti antigen akhalebe pakhungu la nitrocellulose zafotokozedwa mwatsatanetsatane pamwambapa motero zimafotokoza zomwe zili papepalalo.
Zothandizira
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