Pulatifomu Yopanga Aptamer

Ma aptamers ndi oligonucleotide yokhala ndi chingwe chimodzi (DNA, RNA kapena XNA) yokhala ndi mphamvu ya kuyanjana kwakukulu komanso kudziwika bwino komwe kumalumikizana makamaka ndi mamolekyulu monga ma antibodies, ndipo amagwiritsidwa ntchito kwambiri popanga ma biosensors, matenda ndi mankhwala.

Pulatifomu ya aptamer yoperekedwa ndi Alpha Lifetech ili ndi magulu awiri: nsanja yopangira aptamer, yomwe makamaka imakhudza ntchito yopangira aptamer library ya SELEX ndi ntchito yopangira aptamer (DNA, RNA kapena XNA), ndi nsanja yowunikira aptamer kuphatikiza ntchito zowunikira kutengera ukadaulo wa SELEX wa mapuloteni, ma peptide, maselo, mamolekyu ang'onoang'ono, ma ayoni achitsulo ndi mamolekyu ena ofunikira, komanso ntchito zowunikira ndi kusanthula aptamer.

Nsanja Yopangira Aptamer

Utumiki wopangira laibulale ya SELEX aptamer

Ntchito yopangira laibulale ya SELEX aptamer imafuna kwambiri popanga laibulale yokhala ndi mitundu yambiri ya oligonucleotide yokhala ndi chingwe chimodzi mwa kupanga mankhwala mu vitro motsatira mamolekyu omwe akufuna. Kupanga laibulale ndiye poyambira pa ukadaulo wa SELEX, womwe umapereka njira zambiri zowunikira pambuyo pake popanga malaibulale akuluakulu osasinthika ndikuwonjezera mwayi wowunikira ma aptamers okhala ndi mgwirizano wapamwamba.

Kupanga laibulale kumagawidwa m'magawo otsatirawa:

Masitepe	Tsatanetsatane wa Ukadaulo
Dziwani Mamolekyulu Omwe Mukufuna Kudziwa	Dziwani mamolekyu omwe akufunika kufufuzidwa kuti apeze ma aptamers, omwe angakhale mapuloteni, ma nucleic acid, ma molekyulu ang'onoang'ono, ma ayoni achitsulo, ndi zina zotero.
Kapangidwe ka Mndandanda Wosasinthika	Kutalika kwa mndandanda wosasinthika, kapangidwe ka maziko ndi magawo ena adapangidwa malinga ndi mawonekedwe a mamolekyu omwe akufuna komanso zofunikira pakuwunikira. Kawirikawiri, mndandanda wosasinthika umakhala pakati pa makumi ndi mazana a maziko kutalika.
Kupanga kwa Ma Fixed Sequences	Zidutswa za oligonucleotide zokhala ndi ma sequence okhazikika (monga ma PCR primer sequences) kumapeto onse awiri zimapangidwa ndikupangidwa, zomwe zidzagwiritsidwa ntchito pakukulitsa ndikuwunika zotsatira.

Laibulale yopangidwa ikufunikabe kukonzedwanso kuti iwongolere khalidwe. Kuchuluka kwa laibulale kunatsimikizika kuti kutsimikizire kuti ikugwiritsidwa ntchito poyesa zotsatira. Kusiyanasiyana ndi kulondola kwa zochitika zosasinthika mu laibulale kunatsimikiziridwa mwa kutsatana ndi njira zina kuti zitsimikizire kuti khalidwe la laibulale likukwaniritsa zofunikira zoyesa.

Kudzera mu masitepe omwe ali pamwambapa, laibulale ya SELEX aptamer yapamwamba komanso yosiyanasiyana ikhoza kupangidwa, yomwe ingapereke njira zambiri zowunikira zotsatira.

Ntchito zopanga ma Aptamer (DNA, RNA kapena XNA)

Aptamers nthawi zambiri amatanthauza ma aptamers a nucleic acid. Ma aptamers a nucleic acid akuphatikizapo ma aptamers a DNA, ma aptamers a RNA, ndi ma aptamers a XNA omwe ndi ma aptamers a nucleic acid osinthidwa ndi mankhwala. Njira ya SELEX imagwiritsidwa ntchito kwambiri popanga ma aptamers. Njira yoyambira yopangira ma aptamer imaphatikizapo kumanga laibulale, kumangirira zolinga, kudzipatula ndi kuyeretsa, kukulitsa, kufufuza maulendo angapo, ndi kuzindikira mndandanda. Kwa zaka zambiri, takhala tikuyang'ana kwambiri pakupanga laibulale komanso chidziwitso chochuluka pakupanga ma aptamer. Tadzipereka kupatsa makasitomala ntchito zabwino.

Njira ya Ukadaulo ya SELEX

Njira ya SELEX imakhala ndi maulendo angapo, omwe ali ndi magawo ofunikira awa:

Kumanga Laibulale ndi Cholinga

Laibulale ya nucleic acid yomangidwa imasakanizidwa ndi mamolekyulu enaake (monga mapuloteni, mankhwala ang'onoang'ono a mamolekyulu, ndi zina zotero), kotero kuti ma nucleic acid omwe ali mu laibulale akhale ndi mwayi wolumikizana ndi mamolekyulu omwe akufuna.

Kupatula Mamolekyu Osamangidwa

Ma nucleic acid sequences omwe sali omangika ku molekyulu yomwe akufuna kuigwiritsa ntchito amalekanitsidwa ndi chisakanizocho pogwiritsa ntchito njira zinazake monga affinity chromatography, magnetic bead leating, ndi zina zotero.

Kuwonjezeka kwa Mamolekyulu Omangirira

Dongosolo la nucleic acid lomangiriridwa ku molekyulu yomwe mukufuna limakulitsidwa, nthawi zambiri pogwiritsa ntchito ukadaulo wa polymerase chain reaction (PCR). Pa gawo lotsatira lowunikira, magawo okulitsidwa adzagwiritsidwa ntchito ngati laibulale yoyambira.

Chithunzi 1: Njira yowunikira ya SELEX

Nsanja Yowunikira ya Aptamer

Utumiki wowunikira wa Aptamer

Alpha Lifetech imapereka mitundu yosiyanasiyana ya ntchito zapadera zowunikira aptamer pogwiritsa ntchito njira zosiyanasiyana za SELEX pamitundu yosiyanasiyana ya mamolekyu anu:

Mitundu ya Zolinga	Tsatanetsatane waukadaulo
Kuwunika kwa Mapuloteni Aptamer ndi SELEX	Cholinga chachikulu cha kufufuza mapuloteni a aptamer ndikuwunikira ma aptamer omwe amatha kulumikizana mwachindunji ndi mamolekyu a mapuloteni. Ma aptamer awa ndi osavuta kupanga, okhazikika komanso osakhudzidwa ndi zinthu zachilengedwe.
Kuwunika kwa Peptide Aptamer ndi SELEX	Ma aptamers a Peptide ndi gulu la ma peptide afupiafupi okhala ndi mawonekedwe apadera komanso kuyanjana kwakukulu, komwe kumatha kumangirirana mwachindunji ndi zinthu zomwe zimafunidwa ndikuwonetsa kuthekera kosiyanasiyana kwa kugwiritsidwa ntchito m'munda wa biomedical. Kudzera mu njira yowunikira yeniyeni, ma aptamers a peptide omwe amatha kumangirirana mwachindunji ndi mamolekyulu omwe amafunidwa amafufuzidwa kuchokera ku malaibulale ambiri a peptide omwe amasankhidwa mwachisawawa.
Kuwunika kwa Aptamer ya Selo (Cell-SELEX)	Maselo ofunikira kapena mamolekyu enaake pamwamba pa selo amakonzedwa ngati malekyulu. Malekyulu akhoza kukhala maselo athunthu, ma receptor pa nembanemba ya selo, mapuloteni, kapena mamolekyu ena ang'onoang'ono.
Kuwunika kwa Aptamer ya Molekyulu Yaing'ono ndi Capture SELEX	Capture SELEX ndi njira yowunikira ma aptamer ang'onoang'ono a mamolekyulu, yomwe ndi mtundu wa SELEX. Capture SELEX ndi yoyenera kwambiri powunikira ma aptamer ang'onoang'ono a mamolekyulu, omwe nthawi zambiri amakhala ndi magulu ochepa ogwira ntchito ndipo ndi ovuta kuwaletsa mwachindunji pazithandizo zolimba.
Ntchito za SELEX zochokera ku Zinyama Zamoyo	Utumiki wowunikira nyama zamoyo ndi njira yoyesera yomwe imagwiritsidwa ntchito kwambiri m'magawo a sayansi ya zamoyo, zamankhwala ndi ukadaulo wa zamoyo, yomwe imagwiritsa ntchito nyama zamoyo ngati zitsanzo zoyesera kuti ifufuze ndikuwunika mamolekyulu enaake, madokotala, machiritso kapena njira zamoyo. Mautumikiwa adapangidwa kuti ayerekezere malo a thupi m'thupi la munthu kuti alosere molondola ndikuwunika momwe zotsatira za mayeso zimagwirira ntchito komanso chitetezo cha zotsatira za mayeso m'thupi la munthu.

Utumiki wokonza bwino Aptamer

Kukonda madzi, kutayika kwakukulu kwa mphamvu ya amplifice popanga, komanso kutulutsa mwachangu ma amplifice kumalepheretsa kugwiritsa ntchito. Pakadali pano, njira zosiyanasiyana zowongolera zafufuzidwa kuti ziwongolere magwiridwe antchito a amplifice.

Tilinso ndi njira zosiyanasiyana zokonzerera aptamer zomwe zimaphatikizapo kudula, kusintha, kulumikiza ku gulu loyenera (thiol, carboxy, amine, fluorophore, ndi zina zotero).

Utumiki wosanthula zizindikiro za Aptamer

Utumiki wosanthula mawonekedwe a Aptamer umatanthauza ntchito yaukadaulo yowunikira kapangidwe ka magwiridwe antchito ndi kutsimikizira magwiridwe antchito a aptamer yomwe yapezeka kuti iwonetsetse kuti aptamer ikukwaniritsa zofunikira zenizeni zomangirira, kukhazikika ndi kutsimikizika. Makamaka imaphatikizapo kusanthula kwa kuyanjana ndi kutsimikizika, kuwunika kukhazikika ndi kutsimikizira ntchito yachilengedwe.