Zomwe Zimapangitsa Kuti Ma antibodies Apitirize Kugwiritsa Ntchito Nitrocellulose Kakhungu Kotsatira Kuyenda Kwawo?
Nitrocellulose Membrane mu Lateral Flow Assay imabwera pambuyo pa chitsanzo ndi conjugate pad. Nitrocellulose membrane imatchedwanso substrate membrane ndipo ndi imodzi mwa zigawo zofunika kwambiri komanso zomvera kwambiri zotsatira za mayeso omwe amachitika pa Lateral Flow Assay . Mu Lateral Flow Assay, pakhoza kukhala zinthu zosiyanasiyana zomwe zimagwiritsidwa ntchito monga cellulose acetate, polyvinylidene fluoride (PVDF), charge modified nylon, ndi polyethersulfone (PES), ndi zina zotero. Mu Nitrocellulose membrane, pali kukula kwa ma pore, porosity, makulidwe, ndi mankhwala osiyanasiyana . Nitrocellulose membranes siili ndi magawidwe ofanana a ma pores kotero nthawi ya capillary flow ndi yabwino kwambiri. Musanapite patsogolo pa chifukwa chake ndikofunikira kudziwa za kayendedwe ka capillary. Kwenikweni, nthawi ya capillary flow imayesedwa mu masekondi kapena masentimita ndipo ndi nthawi yonse yomwe chitsanzocho chinatenga kuti chidzazidwe kwathunthu mu test strip 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 ya nitrocellulose , ma antibodies amakhala ndi kulumikizana ndi tinthu tachilengedwe kaya latex kapena tinthu tagolide. Ma antibodies saloledwa kuyenda ndipo amalepheretsedwa pa nembanemba ya nitrocellulose chifukwa cha zifukwa zosiyanasiyana. Popeza pali mizere yoyesera ndi mizere yowongolera pa kuyesa kwa lateral flow, ma antibodies amamatira pakati pa mizere iwiriyi. Poganizira mzere woyesera, umagwiritsidwa ntchito kumangirira puloteni ya chitsanzo pomwe mzere wowongolera uli ndi ma antibodies enieni a mtundu womwe amagwiritsidwa ntchito kuzindikira ma antibodies mu chitsanzo. Chifukwa china chomwe ma antibodies amakhala pa nembanemba ya nitrocellulose ndi kukula kwa pores ya filimu yobwerera. Filimu yobwerera kumbuyo yopanda matope ya nembanemba ya nitrocellulose simalola ma antibodies kutsika m'malo mwake amagwiridwa mwamphamvu motero amakhala pa nembanemba. Izi zimathandizanso kuchepetsa kusinthasintha.
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
Pomaliza ndi kufotokozera mwachidule, nitrocellulose ndi gawo lofunika kwambiri la mayeso oyenda mbali ndi chipangizo chomwe chimagwiritsidwa ntchito poyesa. Chimodzi mwazinthu zofunika kwambiri komanso zofunika kwambiri zomwe zimatsimikiza mayeso oyenda mbali ndi kukhudzidwa kwake ndi Substrate Membrane kapena Nitcocellulose Membrane . Mizere yoyesera ndi yowongolera imakokedwa pa nembanemba kuti zitsimikizire kuti ma antibodies amalumikizana bwino komanso kuti agwire bwino. Kusakhazikika kwa ma antibodies pamizere yoyesera ndi yowongolera kumatha kukhudza kukhudzidwa kwa zotsatira, zomwe zimapangitsa kuti zotsatira zake zikhale zabodza. Popeza nitrocellulose ndi chimodzi mwa zigawo za mayeso oyenda mbali zomwe chitsanzocho chimadutsa. Kuchokera ku chitsanzo, ma antibodies ndi magawo a antigenic a chitsanzo ndi chipangizocho amalumikizana wina ndi mnzake ndikupitilira ku gawo lotsatira. Kuchokera pamenepo, chitsanzocho chimapita ku nembanemba ya nitrocellulose yomwe imatenga ma antibodies ndi ma antigen ofunikira poyesa. Katundu ndi zifukwa zomwe zimapangitsa kuti antigen ikhalebe pa nembanemba ya nitrocellulose zafotokozedwa mwatsatanetsatane pamwambapa motero zikukhudza nkhani ya pepalalo.
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