Puratifomu Yekuvandudza Aptamer

MaAptamers imhando ye oligonucleotide ine tambo imwe chete (DNA, RNA kana XNA) ine hunhu hwekubatana kwakanyanya uye hunyanzvi hwakanyanya hunobatana nemamorekuru akanangana nemasero emuviri, uye anoshandiswa zvakanyanya mukugadzira biosensors, kuongororwa uye kurapwa.

Chikuva cheaptamer chinopihwa neAlpha Lifetech chinosanganisira mapoka maviri: chikuva cheaptamer synthesis, icho chinonyanya kusanganisira SELEX aptamer library synthesis service uye aptamer (DNA, RNA kana XNA) development service, uye chikuva cheaptamer screening chinosanganisira masevhisi ekuongorora akavakirwa patekinoroji yeSELEX yemapuroteni, mapeptide, masero, mamorekuru madiki, ioni dzesimbi nedzimwe target molecules, pamwe chete nemabasa ekuongorora eaptamer optimization ne identification.

Puratifomu yeAptamer Synthesis

Sevhisi yekugadzira raibhurari yeSELEX aptamer

Basa rekugadzira raibhurari reSELEX aptamer rinonyanya kusanganisira kuvaka raibhurari ine huwandu hwakawanda hwema "oligonucleotide sequences" asina kurongeka ne "in vitro chemical synthesis" zvichienderana nemamorekuru akatarwa. Kuvakwa kweraibhurari ndiko kutanga kwetekinoroji yeSELEX, iyo inopa ma "candidate sequences" akawanda ekuongorora anotevera nekuvaka maraibhurari makuru asina kurongeka uye inowedzera mukana wekuongorora ma "high-affinity aptamers".

Kugadzira raibhurari kwakakamurwa kuita matanho anotevera:

Matanho	Ruzivo rweTekinoroji
Ziva Mamorekuru Aunoda	Ziva mamorekuru anofanirwa kuongororwa kuti aonekwe seane maaptamers, anogona kunge ari mapuroteni, nucleic acids, mamorekuru madiki, maion esimbi, nezvimwewo.
Dhizaini Yekutevedzana Kwenguva Isina Kurongwa	Kureba kwezvikamu zvisingatarisirwi, kuumbwa kwechigadziko nezvimwe zvinhu zvakagadzirwa zvichienderana nehunhu hwemamorekuru anotarisirwa uye zvinodiwa pakuongorora. Kazhinji, zvikamu zvisingatarisirwi zvinosvika pakati pemakumi nemazana ezvikamu.
Kugadzirwa kweFixed Sequences	Zvidimbu zveoligonucleotide zvine ma sequences akagadzika (akadai sePCR primer sequences) kumativi ese akagadzirwa uye akagadzirwa, ayo achashandiswa mukuwedzera nekuongorora zvinotevera.

Raibhurari yakagadzirwa ichiri kuda kugadziriswa zvakanyanya kuti ionekwe kuti mhando yeraibhurari yacho ndeyei. Kuwanda kweraibhurari kwakaonekwa kuti kunoshanda here mukuongorora kwakatevera. Kusiyana-siyana uye kururama kwezvikamu zvisina kurongeka muraibhurari kwakasimbiswa nekuongorora nekutevedzana nedzimwe nzira dzekuona kuti mhando yeraibhurari yaizadzisa zvinodiwa zvekuongorora.

Kuburikidza nematanho ari pamusoro apa, raibhurari yeSELEX aptamer yemhando yepamusoro uye yakasiyana-siyana inogona kugadzirwa, iyo inogona kupa akawanda marongero ekuongorora anotevera.

Masevhisi ekugadzira Aptamer (DNA, RNA kana XNA)

Aptamers dzinowanzoreva ma aptamers e nucleic acid. Ma aptamers e nucleic acid anosanganisira ma aptamers e DNA, ma aptamers e RNA, uye ma aptamers e XNA ayo anogadziriswa nemakemikari e nucleic acid aptamers. Nzira yeSELEX inoshandiswa zvakanyanya pakugadzira ma aptamers. Mafambiro ekutanga emabasa ekugadzira ma aptamer anosanganisira kuvaka raibhurari, kusunga chinangwa, kupatsanura nekucheneswa, kuwedzera, kuongorora kakawanda, uye kuziva sequence. Kwemakore mazhinji, tave tichitarisa pakuvaka raibhurari uye ruzivo rwakakura mukugadzira ma aptamer. Takazvipira kupa vatengi masevhisi ari nani.

Maitiro eTekinoroji yeSELEX

Maitiro eSELEX ane madenderedzwa akawanda, rimwe nerimwe rine matanho akakosha anotevera:

Kubatanidza Raibhurari neTarget

Raibhurari ye nucleic acid yakavakwa inosanganiswa nemamorekuru chaiwo (akadai semapuroteni, mamorekuru madiki, nezvimwewo), kuitira kuti manucleic acid sequences ari muraibhurari ave nemukana wekusungana nemamorekuru acho.

Kuparadzaniswa kweMamorekuru Asina Kusungwa

Nucleic acid sequences dzisina kusungirirwa kune target molecule dzinoparadzaniswa kubva mumusanganiswa nenzira dzakatarwa dzakadai se affinity chromatography, magnetic bead separation, nezvimwewo.

Kuwedzera kweMamorekuru Anosunga

Kutevedzana kwe nucleic acid kunobatanidzwa nemorekuru inotarirwa kunowedzerwa, kazhinji uchishandisa tekinoroji yepolymerase chain reaction (PCR). Pachikamu chinotevera chekuongorora, kutevedzana kwakawedzerwa kuchashandiswa seraibhurari yekutanga.

Mufananidzo 1: Maitiro ekuongorora SELEX

Chikuva Chekuongorora Aptamer

Sevhisi yekuongorora Aptamer

Alpha Lifetech inopa masevhisi akasiyana-siyana ekuongorora aptamer achishandisa nzira dzakasiyana dzeSELEX dzemhando dzakasiyana dzemamorekuru ako:

Mhando dzezvinangwa	Ruzivo rwehunyanzvi
Kuongorora Mapuroteni Aptamer neSELEX	Chinangwa chikuru chekuongorora mapuroteni aptamer ndechekuongorora maaptamer anogona kubatana nemamorekuru eprotein akananga. Maaptamer aya ari nyore kugadzira, akasimba uye haanyanyi kukanganiswa nezvinhu zvezvakatipoteredza.
Kuongororwa kwePeptide Aptamer neSELEX	Ma aptamers ePeptide iboka rema peptide sequences mapfupi ane hunhu hwakanyanya uye hukama, ayo anogona kubatana zvakananga nezvinhu zvinotarwa uye kuratidza mukana wakakura wekushandiswa mumunda we biomedical. Kuburikidza nemaitiro chaiwo ekuongorora, ma aptamers epeptide anogona kubatana zvakananga nemamorekuru anotarwa anoongororwa kubva mumaraibhurari akawanda e random peptide sequence.
Kuongorora Aptamer Yesero (Cell-SELEX)	Masero akagadzirirwa kurongeka kana mamorekuru chaiwo ari pamusoro pesero anogadzirwa sematarget. Matarget anogona kuva masero akazara, mareceptor ari pamembrane yesero, mapuroteni, kana mamwe mamorekuru madiki.
Kuongororwa kweAptamer yeMorekuru Diki neCapture SELEX	Capture SELEX inzira yekuongorora maaptamer madiki emamorekuru, inova imwe mhando yeSELEX. Capture SELEX inonyanya kukodzera kuongorora maaptamer ezvinangwa zvemamorekuru madiki, ayo anowanzova nemapoka mashoma anoshanda uye akaoma kumisa kufamba zvakananga pazvitsigiro zvakasimba.
Masevhisi eSELEX anowanikwa paMhuka Mhenyu	Basa rekuongorora mhuka mhenyu inzira yekuedza inoshandiswa zvakanyanya muminda yesainzi yezvesainzi, mushonga uye tekinoroji yezvesainzi, iyo inoshandisa mhuka mhenyu semhando dzekuedza kuongorora nekuongorora mamorekuru chaiwo, chiremba, kurapa kana maitiro ezvesainzi. Mabasa aya akagadzirirwa kutevedzera nharaunda yezvesainzi mumuviri wemunhu kuti ifanotaura uye iongorore kushanda uye kuchengetedzeka kwemhedzisiro yekuedza mumuviri wemunhu.

Sevhisi yekugadzirisa Aptamer

Kunyungudika kwemvura, kurasikirwa kwakanyanya kweaffinity panguva yekugadzira, uye kukurumidza kubviswa kwema aptamers kunodzikisira kushandiswa kwawo. Parizvino, nzira dzakasiyana-siyana dzekugadzirisa dzakaongororwa kuti dziwedzere kushanda kwema aptamers.

Tinewo nzira dzakasiyana-siyana dzekugadzirisa aptamer dzinosanganisira kudimbura, kugadzirisa, kubatana neboka rakakodzera (thiol, carboxy, amine, fluorophore, nezvimwewo).

Basa rekuongorora hunhu hweAptamer

Basa rekuongorora hunhu hweAptamer rinoreva basa rehunyanzvi rekugadzirisa maitiro ekuongorora mashandiro uye kusimbisa mashandiro eaptamer yakawanikwa kuona kuti aptamer inosangana nezvinodiwa zvekusunga, kugadzikana uye hunyanzvi. Inonyanya kusanganisira kuongorora hukama uye hunyanzvi, kuongorora kugadzikana uye kusimbisa mashandiro emuviri.