Basa reKukura kweAntibody Affinity

Alpha Lifetech Inc.tinogona kupa sevhisi yekukura kweaffinity yeantibody nekushandisa matekinoroji ekuchinja uye kusarudza. Tinowanzo shandisa scFv sefomati yeaantibody mukuita kweaffinity maturation service uye monovalent display phagemid system iyo yakashandiswa kuderedza mhedzisiro yeavidity panguva yekuongorora antigen-binding. Chikwata chedu chehunyanzvi chinogona kupa masevhisi ekukura kweaffinity ye single-domain antibodies zvakare.

Alpha Lifetech Inc.inodada nekupa mabasa akazara kuti isangane nezvinodiwa zvakasiyana-siyana zvevatengi vepasi rose. Chinangwa chedu ndechekunzwisisa nekusangana nezvinodiwa nevatengi vakasiyana uye kubatsira nematambudziko ari kuuya uye ari kubuda mubasa rekutsvagisa.

Chii chinonzi Kukura Kwehukama?

Kukura kweAffinity inzira iyo ma antibodies anodzokororwa kuchinja uye kusarudzwa kuti awedzere kubatana kwawo kune maantigen chaiwo. Masero eB anonyanya kuitika pakati pema germinal organs e secondary lymphoid, anozopedzisira aita somatic hypermutation uye kusarudzwa pasi pesimba reT cells ne follicular dendritic cells.
Munguva yekukura kweaffinity, masero eB ane low-affinity surface immunoglobulin receptors anoshandiswa ne antigen kusangana uye anowedzera kuwanda nekusiyana. Kuburikidza ne somatic hypermutation affinity maturation, random nucleotide substitutions inoitika munzvimbo inoshanduka yemajini e immunoglobulin, zvichikonzera kugadzirwa kwemarudzi akasiyana-siyana e antibody. Mushure me somatic hypermutation, sarudzo inotungamirwa ne antigen inoitwa pamasero eB ane mutated immunoglobulin receptors. Masero eB anogadzira ma antibodies ane antigen affinity yakawedzerwa anochengetwa zviri nani uye anobvumirwa kuwanda, nepo masero eB ane affinity yakaderera kana isina kuchinja anochengetwa apoptosis. Iyi nzira yekudzokorora yekuchinja nekusarudza inotungamira mukugadzirwa kwemasero eB, ichigadzira ma antibodies ane affinity yakakwira zvishoma nezvishoma ye antigen.

Basa reKukura kweAntibody Affinity

Zvakanakira zvikuru zvePCR-based methods ndezvekuti mutations dzakanangana zvakananga ne amplified fragment; error rate iri nyore kudzora uye nzira yacho inokurumidza uye iri nyore kuisa uye haishandisi makemikari ane njodzi. Zvinozivikanwa kuti Taq DNA polymerase inodzokorora DNA zvisina kusimba, zvakanyanya nekuda kwekushaikwa kwe3' kusvika 5' process yekuongorora.Alpha Lifetech'sChikuva chekugadzira maantibody chinoshandisa nzira yePCR inokanganisa mafambiro ekuchinja nzvimbo dzeCDR panguva yekuvaka raibhurari iri pasi pema "sub-raibrary". Uye kazhinji tinogadzira shanduko munzvimbo dzisina kurongeka muzvikamu zvese zveVH neVL, kuitira kuwedzera kusiyana kwemajini e "sub-raibrary". Mushure mekuvakwa kweraibhurari yemajini e "antibody" yakachinja nePCR inokanganisa mafambiro e "error-prone", kusarudzwa kwemhando dzakasiyana dze "high-affinity variants" (kubatana kwema antibodies e-scFv kunogona kusvika 10^8 - 10^10 M) kunoitwa nekuisa mu "panning" kana pa "immobilized antigen" ine mamiriro ekugezesa akagadzirirwa kusarudzwa kusingatarisirwi.
  
Masayendisiti muAlpha Lifetech Inc.shandisai Escherichia coli mutator strains mutAD5TM seimwe yenzira dzakasiyana-siyana dzekuchinja kuti muunze shanduko dzisina kurongeka, uye nokudaro mushandure hukama uye kutaura kwezvikamu zve recombinant antibody. Mamiriro ekusarudza anogona kuchinjwa kune zvikamu zve antibody zvine huwandu hwakawanda hwekugadzirwa. Mamiriro ekukura mumasero eEscherichia coli mutator anogona kugadziriswa kuti aunze shanduko imwe chete yepfungwa isina kurongeka pa kilobase yeDNA, inenge yakaenzana nekuchinja kwecodon imwe pa scFv fragment. Mushure mematanho akati wandei ekuchinja, kuratidzwa, uye kusarudzwa, tinogona kuvandudza hukama hwehukama hwekubva pa10^5 - 10^6 M kusvika 10^8 - 10^10 M.

Kuvandudza Maitiro Ekukura Kwehukama

Dzimwe nzvimbo dzema antibodies dzinogona kurongwa nenzira dzakasiyana-siyana (sekunge kurongwa kwakazara nema amino acid ese makumi maviri kana kurongwa kwakasarudzika nema amino acids akasarudzwa pahuwandu hwakatarwa) kuti tivandudze hukama. Tine nzira mbiri dzekuvandudza hukama hwema antibody: chekutanga, shanduko dzinounzwa panzvimbo dzakaganhurirwa munzvimbo dzinosarudza kubatana (CDRs) nekuchinja kwakanangana nenzvimbo; chechipiri, shanduko dzinounzwa munzvimbo dzese dzeV-coding nekuchinja kwakasarudzika. Nekushandisa nzira idzi mbiri, ma amino-acid akachengetwa kana nzvimbo yese ye framework ye antibody ichatsiviwa nemamwe ma amino acids, ayo akabatanidzwa ne peptide library scanning technologies edu, anogona kuwedzera hukama hwema antibodies anodiwa kusvika padanho repamusoro.

Kubatana kwePhage Display-Antibody

Kana raibhurari ye scFv mutant yagadzirwa, nzira mbiri dzekuongorora dzinowanikwa: biopanning uye solid-antigen sorting. Pakutanga, huwandu hwe substrate hunoshandiswa kupatsanura high-affinity antibody, sezvo low-affinity mutants ichasukwa, zvichisiya high-avidity phage particles. Nzira yechipiri inoshandisa antigen yakanyorwa mu solution, selection yakavakirwa pa equilibrium constant (Kd), uye selection yakavakirwa pa binding kinetics, nzira iyi yekusarudza inosiyanisa antibody variants ine improved Kd.

Kana muine mibvunzo, ndapota inzwa wakasununguka kutibata chero nguva.