Ntev R3 IGF-I(link:https://www.bloomtechz.com/synthetic-chemical/peptide/long-r3-igf-i-cas-143045-27-6.html) yog ib qho hluavtaws polypeptide molecule uas nws tshawb pom keeb kwm pib hauv 1970s. Lub sijhawm ntawd, cov kws tshawb fawb tau pib ua tib zoo saib lub luag haujlwm tseem ceeb ntawm endogenous insulin-zoo li kev loj hlob yam-I (IGF-I) hauv kev tswj kev loj hlob thiab cov metabolism, thiab sim tsim cov qauv molecular zoo ib yam li IGF-I tab sis ntau yam tshuaj lom neeg thiab tshuaj. Ib hom tshiab ntawm peptide molecule nrog daim ntawv thov nqi.
1. Kev tshawb nrhiav thiab kev tshawb fawb ntawm IGF-I:
Thaum ntxov xyoo 1950, cov kws tshawb fawb tau pib tshawb nrhiav qhov muaj thiab kev ua haujlwm ntawm kev loj hlob zoo li insulin. Hauv xyoo 1960, qee lub koom haum tshawb fawb tau cais tawm ib hom tshiab ntawm cov protein nrog kev loj hlob ntawm tes thiab txhawb kev loj hlob los ntawm tsiaj cov ntshav, hu ua kev loj hlob hormone (GH). Tom qab ntawd, cov kws tshawb fawb tau tshawb pom lwm cov protein uas ze ze rau GH los ntawm cov tsiaj cov ntshav thiab lwm cov ntaub so ntswg, hu ua IGF-I.
IGF-I yog cov protein me me uas muaj 70 amino acid residues, thiab nws cov qauv zoo ib yam li tib neeg insulin. IGF-I feem ntau yog tsim los ntawm lub siab, uas yog ze ze rau lub physiological teebmeem ntawm GH, thiab tuaj yeem tswj cov cell proliferation, sib txawv thiab cov metabolism hauv kev sib cuam tshuam ntawm nws tus kheej receptors thiab insulin zoo li kev loj hlob receptor (IGF-IR).
Hauv xyoo 1970, raws li kev tshawb fawb ntawm IGF-I tob zuj zus, cov kws tshawb fawb tau pib tshawb nrhiav nws cov qauv molecular thiab cov khoom siv roj ntsha, thiab sim tsim kom muaj txiaj ntsig zoo dua IGF-I analog molecule.
2. Tshawb nrhiav thiab tshawb fawb txog ntev R3 IGF-I:
Los ntawm lig 1970s mus rau thaum ntxov 1980s, qee cov kws tshawb fawb tau pib hloov N-terminal sib lawv liag ntawm IGF-I thiab tsim ib qho IGF-I analog nrog cov qauv molecular ruaj khov thiab yooj yim synthesis thiab siv. Raws li qhov no, ntev R3 IGF-Kuv tau yug los.
Ntev R3 IGF-I siv arabinosyl-Ala-Pro-Ala (Apa) los hloov Gln-Pro-Arg-Gly ib ntus ntawm endogenous IGF-I, ua rau lub neej ntev dua ib nrab ntawm cov ntshav, thiab tsis yooj yim khi thiab tshem tawm los ntawm IGF-binding protein (IGFBP). Tsis tas li ntawd, ntev R3 IGF-I kuj tau hloov kho los ntawm kev ntxiv 13 amino acid sequences (xws li Arg-Lys-Glu-Gly-Ser) ntawm C-terminus, qhia disulfide bonds thiab -helical qauv, thiab lwm yam, kom nws muaj kev ua haujlwm lom neeg ntau dua thiab muaj peev xwm rau kev siv tshuaj.
Thaum lub sij hawm kev tshawb fawb thiab kev loj hlob ntawm ntev R3 IGF-I, qee cov kws tshawb fawb kuj tau sim los txhim kho nws cov kev ua tau zoo thiab cov nqi tsim khoom los ntawm kev siv tshuab transgenic thiab lwm yam. Piv txwv li, ntev R3 IGF-I tau qhia los ntawm microbial systems xws li Escherichia coli thiab poov xab, thiab purified thiab sib cais los ntawm acid kho, countercurrent chromatography thiab lwm yam technologies, thiab thaum kawg ib high-purity ntev R3 IGF-I khoom tau txais.
Thaum lub sijhawm kev tshawb fawb ntev, raws li cov qauv tshwj xeeb ntawm LONG R3 IGF-I, uas yog polypeptide molecule zoo ib yam li cov qauv rau endogenous IGF-I thiab muaj 13 amino acids ntxiv, ntau yam khoom siv hluavtaws tau kawm rau kev tsim khoom. Cov txheej txheem kev npaj ntawm ntev R3 IGF-I feem ntau muaj cov hauv qab no:
1. Chemical synthesis method:
Tshuaj synthesis yog ib qho ntawm feem ntau siv txoj kev npaj rau ntev R3 IGF-I. Cov tshuaj synthesis ntawm ntev R3 IGF-I tau ua raws li lub npe hu ua amino acid sequence ntawm IGF-I, thiab ntxiv 13 amino acid sequences ntxiv ntawm N-terminus ntev R3 IGF-I. Synthesis yuav tsum tau siv ntau pab pawg tiv thaiv kom paub meej cov amino acid selectivity thiab cov tshuaj tiv thaiv efficiency. Feem ntau, cov kev tiv thaiv peptide ntu ntawm lub hom phiaj amino acid yog thawj zaug npaj los ntawm cov khoom siv-theem synthesis, thiab tom qab ntawd sib sau ua ke rau hauv R3 IGF-I molecule ntev los ntawm cov kua-theem synthesis.
2. Biotechnology Act:
Cov txheej txheem biotechnology tsuas yog siv cov hlwb tsim los nthuav qhia cov proteins sib txuas, thiab nthuav tawm LONG R3 IGF-I los ntawm kev hloov cov noob kab ke thiab kev qhia vectors. Hauv cov qauv no, LONG R3 IGF-I noob tuaj yeem nkag mus rau hauv lub xov tooj ntawm tes rau kev qhia los ntawm gene recombination technology, lentiviral vector, plasmid vector thiab lwm yam. Txoj kev no tuaj yeem tsim qhov loj ntawm LONG R3 IGF-I, thiab tuaj yeem ua kom zoo dua nws cov kev qhia thiab kev ua kom huv huv los ntawm kev hloov cov vector thiab cov teeb liab zais cia.
3. Txoj kev Enzymatic:
Txoj kev enzymatic tsuas yog siv cov enzymes tshwj xeeb xws li pepsin thiab clam nqaij enzyme kom tshem tawm qhov ntev R3 IGF-I precursor protein kom tau txais LONG R3 IGF-I monomer, thaum zam kev tsis tsim nyog los ntawm cov khoom lag luam. Nyob rau hauv txoj kev no, cov matrix uas muaj qhov ntev R3 IGF-I precursor protein yuav tsum tau ua ntej, thiab tom qab ntawd reacted ntawm qhov tsim nyog kub los ntawm kev ntxiv enzymes thiab pH tswj, thiab lwm yam, thaum kawg tau lub hom phiaj LONG R3 IGF-I.
4. Protein modification method:
Txoj kev hloov kho cov protein feem ntau yog siv cov synthesized endogenous IGF-I los hloov nws kom ua tiav cov nyhuv ntev R3 IGF-I. Nyob rau hauv txoj kev no, N-terminal ntawm endogenous IGF-I feem ntau yog nkag mus rau hauv 13 cov kab ke tshwj xeeb kom nws muaj cov nyhuv ntev R3 IGF-I. Tsis tas li ntawd, kev ua haujlwm lom neeg thiab ib nrab ntawm lub neej ntev R3 IGF-I tuaj yeem txhim kho ntxiv los ntawm kev hloov pauv pawg C-terminal.
Txhawm rau ua kom tiav, cov txheej txheem synthesis ntawm ntev R3 IGF-I suav nrog cov tshuaj synthesis, biotechnology, enzymatic thiab protein hloov pauv, thiab txhua txoj kev muaj nws qhov zoo, qhov tsis zoo thiab thaj tsam ntawm daim ntawv thov. Nrog rau kev txhim kho txuas ntxiv ntawm kev siv thev naus laus zis thev naus laus zis, genetic engineering thev naus laus zis thiab lwm yam kev lag luam, kev npaj thev naus laus zis ntev R3 IGF-I kuj tseem yuav txhim kho thiab txhim kho ntxiv.