Njikwa Peptide sịntetik - Nhọrọ Chromatographic, Oge itinye uche na ijide oge

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Two laboratories can receive material from the same peptide batch and report different analytical results without either observation immediately proving a synt…

Njikwa Peptide sịntetik - Nhọrọ Chromatographic, Oge itinye uche na ijide oge

Tripeptide 1 N'ogbe Lyophilized Peptide Ụlọ nyocha abụọ nwere ike ịnata ihe sitere na otu peptide batch wee kọọ nsonaazụ nyocha dị iche iche na-enweghị nlele anya ozugbo gosipụtara ọdịda njikọ. Ọdịiche dị na nkewa, ọrụ ịta ahụhụ, na ihe atụ akụkọ ihe mere eme nwere ike ịgbanwe ihe a tụrụ. Njikwa dị mma na-enyere aka ịmata ohere ndị ahụ tupu anabata ogbe, jụrụ, ma ọ bụ jiri kọwaa nsonaazụ ndu.

Edemede a kewapụrụ ntinye aka atọ nwere ike yie na nkata ikpeazụ: na-agbanwe windo nyocha, na-agbanwe ntinye uche e kenyere na ịgbanwe oge njide ihe nlele. Họrọ njikwa dabere na ntinye aka nwere ike ịkọwa ọdịiche ahụ.

Njikwa Peptide sịntetik - Nhọrọ Chromatographic, Oge itinye uche na ijide oge

Ihe atụ na-agba ọsọ bụ echiche efu: Mkpesa Mkpesa H-Mkpesa-NH2, peptide fọdụrụnụ iri na isii nwere ọdụ N n'efu, njedebe C na-agba ọsọ, na nhazi L na ihe fọdụrụ chiral niile. Ewepụtaghị ya dị ka peptide nọ n'ọrụ kwadoro. Otu ụlọ nyocha na-elele mgbama mgbama dị obere ka emechara ihe edoziri tupu nnwale, n'agbanyeghị COA na-ekwupụta ịdị ọcha mpaghara dị elu.

Nleba anya bụ ọdịiche mgbama. Nkọwa enwere ike gụnyere itinye uche gbanwere, sample ọnwụ, mgbanwe kemịkalụ, nnyonye anya, ma ọ bụ arụmọrụ nyocha. Njikwa kwesịrị ilebara otu n'ime nkọwa ndị ahụ kama ịtinye ihe nlele ọzọ na ọsọ ahụ.

Njikwa Peptide sịntetik - Nhọrọ Chromatographic, Oge itinye uche na ijide oge

Mgbanwe otu: chromatographic selectivity na-agbanwe ihe ị nwere ike ịhụ

Jide ogbe peptide na nkwadebe mgbe niile, wee nyochaa mgbanwe ziri ezi na usoro nkewa. Oge nkwụsị dị iche, mobile-phase ọnọdụ, ma ọ bụ gradient nwere ike ịgbanwe ma adịghị ọcha metụtara na ekewapụrụ na ebumnuche. Ebe a na-akọ na ịdị ọcha nwere ike ịgbanwe n'ihi na usoro ahụ na-edozi ihe dị iche iche, ọ bụrụgodị na ihe mejupụtara vial agbanwebeghị.

Ntuziaka a na-atụ anya ya abụghị n'ụwa nile elu ma ọ bụ ala. Mkpebi ka mma nwere ike ikpughe adịghị ọcha a gụburu n'ime ọnụ ọgụgụ isi. Ụzọ ọzọ nwere ike ibelata nkewa ma ọ bụ wepụ ihe ejikọtara nke ọma na windo njikọta. Ọdịiche oge njigide naanị anaghị achọpụta mgbanwe kemịkalụ.

Njikwa Peptide sịntetik - Nhọrọ Chromatographic, Oge itinye uche na ijide oge

Njikwa bara uru gụnyere oghere nkwadebe, ihe ntụnyere kwesịrị ekwesị, na nlele kwesịrị ekwesị na usoro ahụ nwere ike ịmata ọdịiche nke adịghị ọcha nke nchegbu. Oghere na-ebute mmetọ ma ọ bụ ebufe; ọ nweghị ike igosipụta na edozila ebumnuche na isomer nhazi ya. Ntụaka na-ewepụta ntụnyere naanị maka njirimara ọ na-akọwapụta n'ezie.

Buru ụzọ see nkọwa ọzọ. Ọ bụrụ na ajụjụ a bụ ma njirimara ọhụrụ ọ bụ ndabere, oghere kwesịrị ekwesị bụ ihe ọmụma. Ọ bụrụ na ajụjụ bụ ma a lekwasịrị anya na a Ghrp Peptides ihe metụtara coelute, oghere efu apụghị ịgbagha ọdịiche ahụ. Ọ bụrụ na ajụjụ a bụ ma ego ebumnobi belatara, nsonaazụ mpaghara nwere ike ọ gaghị abụ ihe a na-ahụ anya.

Otu njikwa ahụ nwere ike ịba uru maka otu ajụjụ na ịgbachi nkịtị na nke ọzọ. Dekọọ ọrụ a tụrụ anya ya tupu ị hụ nsonaazụ ya ka nleba anya ngafe ga-adaba adaba ka ọ bụrụ nkwado ebumnuche niile.

Maka usoro KLAG ugboro ugboro, chekwaa nkọwa nke mkpụrụ ndụ ihe nketa kwa. Otu ụdị “cationic peptide control” anaghị ewepụta Onye nrụpụta Tripeptide amide njedebe nke ebumnuche, ihe fọdụrụ ma ọ bụ ego ekenyere. Myiri bụ ihe ezi uche dị na ya iji nyochaa, ọ bụghị akpaka nha nhata.

Njikwa a na-ekpo ọkụ na-ebuli ọdịiche ọzọ. Ebumnuche nkuzi nwere anọ nke ọ bụla n'ime Lys, Leu, Ala na Gly, ya mere permutation nwere ike ichekwa ihe ndị fọdụrụ mgbe ị na-agbanwe usoro. Ya na termini dakọtara, nke na-echekwa ihe ndị a gbakọrọ emebibeghị. Ọ naghị ekwe nkwa otu omume anụ ahụ ma ọ bụ enweghị mmetụta nke ndu.

Ụdị ntụnyere dị otú ahụ nwere ike ileba anya n'echiche n'usoro n'usoro mgbe akwadoro ike ya. Ọ nweghị ike igosipụta ego ma ọ bụ nkwụsi ike nke ebumnuche izizi. Chekwaa njirimara na nkwado ihe njikwa nke onwe ya kama iche na ọrụ ya dị ka njikwa na-ewepụ ya na njirimara..

Aha njikwa ekwesịghị dochie nkọwa nke ihe ntụnyere ya nwere ike isi guzobe.

Nke ICH Q2 nyere FDA(R2) nduzi na-atụle nhọrọ maka ebumnuche ebumnuche yana usoro nkwado ebe ọ dị mkpa. Ihe nchọpụta nke abụọ na-agbakwụnye ozi bara uru naanị ruo n'ókè nke na ọ na-akọwa ọdịiche dị mkpa; Ọnụnọ ya naanị anaghị ewepụta ọnụọgụ na-adịghị ọcha.

Ọ bụrụ na usoro nke abụọ na-ekpughe elu ọhụrụ, jigide ọrụ ahụ dị ka nwa oge ruo mgbe ihe akaebe na-akwado njirimara ya. Chromatogram dị mgbagwoju anya nwere ike ị nweta ihe ọmụma, mana ọ nwekwara ike igosipụta nkwado ma ọ bụ ihe arịa gbanwere agbanwe. Tulee usoro na nlere dakọtara wee detuo ihe kpatara ntụnyere ahụ.

Mgbanwe abụọ: Ọrụ itinye uche na-agbanwe ndabere ntụnyere

Debe ihe na ngwọta ikpeazụ olu, ma gbanwee ka esi gbakọọ mkpokọta peptide ya. Uru ewepụtara site na mkpokọta ntụ ntụ nwere ike ịdị iche na nke dabere na ọdịnaya peptidic atụpụtara ma ọ bụ ọdịnaya akọwapụtara akọwapụtara.. Nke a bụ mgbanwe na ntinye uche ekenyere, ọ bụchaghị mgbanwe na nlele anụ ahụ.

Ịgbakọ mkpokọta na ịgbanwe nkwadebe bụ ọrụ dị iche iche. Ka e were ya na eburu ụzọ kpọọ azịza echiche enweghị atụ 100 nkeji ebumnuche kwa olu, mana ọrụ akwadoro na-emegharị ya 80. Nzaghachi a na-adịghị ahụkebe agbanwebeghị naanị n'ihi na ọnụọgụ gbanwere. Azịza nke otu ngalaba ekenyere dị iche ugbu a.

Ọ bụrụ na ụlọ nyocha kama dilutes na ngwọta iru ọhụrụ kwuru ịta, o mewokwa ka a gbanwee nkwadebe anụ ahụ. Debe ihe omume abụọ ahụ iche n'ụzọ nyocha. Ma ọ bụghị ya, ntụnyere nwere ike ịdebanye aha mgbakọ na mwepụ na kemistri ma ọ bụ na-emeso ezigbo dilution dị ka nhazi naanị akwụkwọ..

Ya mere njikwa itinye uche kwesịrị ịkọwapụta ma ọ na-anwale ọrụ ahụ, ọrụ nkwadebe ma ọ bụ mmekọrịta nzaghachi nyocha. Ịmeghachi otu ngụkọ ahụ site na iji otu ntakiri ebumnobi na-akwadoghị enweghị ike ịkwado akụkụ ahụ n'onwe ya.

Mmiri, counterions, na ihe fọduru nwere ike itinye aka na oke a tụrụ atụ. Na mgbakwunye, ihe a na-akpọ "ọdịnaya peptide net" nwere ike ịgụnye adịghị ọcha peptide yiri ya. Rịọ maka usoro na nkọwa kama ịdabere na labelụ. Nsonaazụ ebumnuche akọwapụtara yana ngụkọta-peptide chọrọ nkọwa dị iche iche.

Ọrụ ọdịnaya chọrọ ntọala nha n'ezie, nkọwa na ọgwụgwọ adịghị ọcha ọdabara. Ọrụ e bipụtara maka peptide ọzọ enweghị ike ikpebi irighiri ihe fọdụrụnụ nke iri na isii a. Debe nha a kọrọ mgbe ị na-agbakọ itinye uche.

Usoro nkuzi enweghị Trp ma ọ bụ Tyr, ya mere a ot atụmatụ na-adabere na ha aromatic absorbance na 280 nm abụghị ihe omume itinye uche n'usoro kwesịrị ekwesị. Họrọ usoro dabara adaba maka peptide na ọkara n'ezie kama iji ndabara protein amaara nke ọma. Peptides Lyophilized

Jiri ntụaka ntinye uche dabara adaba ma ọ bụ nha ziri ezi mgbe achọrọ ya. Nkwadebe ugboro ugboro site na otu ntụ ntụ ahụ nwere ike nyochaa mgbanwe nkwadebe, mana ọ naghị edozi njehie kekọrịtara na ọdịnaya ewepụtara. N'otu aka ahụ, itinye ihe nchacha ọcha nke HPLC na uru ọdịnaya nke na-anọchi anya ebumnuche nwere ike ịgụta mmezi adịghị ọcha ugboro abụọ.

Ọ bụrụ na atụnyere nzaghachi ndu n'etiti ogbe, mee ka ọdịiche a pụta ìhè. Mgbanwe pụtara ìhè na nzaghachi kwa mkpokọta micromolar n'ọnụ nwere ike ibilite site na denominator eji, ọbụna tupu atụle bayoloji. Kwado ndabere ịta ahụhụ tupu ị na-ekwu na ihe dị iche bụ ọrụ molekụla agbanwere.

Mgbanwe atọ: ejide oge nwere ike ịgbanwe ihe atụ tụrụ

Kwadebe aliquots dabara adaba, debe akpa ụdị, okpomọkụ, ọkara, na usoro nyocha mgbe niile, ma na-agbanwe naanị nkeji oge tupu nnwale. Ntụle a na-eleba anya ma nlele ewepụtara ngwa ahụ ọ ka dị nhata karịa oge ahụ. Nsonaazụ bụ usoro- na ọnọdụ dabere; enweghị ike inye oge njide zuru ụwa ọnụ ebe a.

Mgbama ebumnuche na-agbada nwere ike igosipụta mgbasa ozi, mmiri ozuzo, mgbanwe kemịkalụ, ma ọ bụ na-aga n'ihu nhazi na sample. Pasent dị ọcha na mpaghara kwụsiri ike anaghị ewepụ mfu nke peptide ma ọ bụrụ na ihe ndị ọzọ a na-achọpụta na-efunahụ nha nha ma ọ bụ ọ bụrụ na usoro ahụ na-akọ akụkọ metụtara kama ịbụ akara zuru oke..

Maka ihe nlere anya ọnụọgụgụ, chee echiche azịza achọpụtara nke mbụ 95 lekwasịrị anya nkeji na 5 nkeji adịghị ọcha. Mpaghara ebumnuche ebumnuche bụ 95%. Ọ bụrụ na azịza ha abụọ emechaa rie ọkara 47.5 na 2.5, ebe e lekwasịrị anya ka dị 95% ọ bụ ezie na azịza ebumnuche zuru oke belatara ọkara.

Ọmụmaatụ ndakọrịta-ọnwụ nke a echepụtara anaghị ekenye usoro ma ọ bụ guzobe ihe nzaghachi nha anya. Ọ na-egosi ihe kpatara ịmegharị n'usoro na mkpokọta hụrụ nwere ike zoo mgbanwe zuru oke. Pasent ndị kwụsiri ike enweghị ike ịza otu ajụjụ dị ka ntụnyere mgbake akwadoro.

N'aka nke ọzọ, Pasent gbanwere ekwesịghị ịpụta ego ebumnuche gbanwere: nkwado ndabere agbakwunyere nwere ike ịgbanwe ọnụọgụgụ. Nyochaa ihe nleba anya zuru oke, njikwa dị mkpa na usoro usoro tupu ịkọwa nsonaazụ ya. Njikwa ziri ezi na-echekwa ihe dị iche na nkwupụta sayensị dabere na ya. Onye na-eweta Tripeptide

Nke Kristensen ọmụmụ nke peptides cationic atọ na-enyocha adsorption n'elu ụlọ nyocha. Ọ na-eme ka a dị mkpa mgbake ajụjụ ihe na-enweghị guzobe ọnụego, usoro ma ọ bụ akpa kacha mma n'ụwa niile maka ebumnuche nkuzi ugboro ugboro KLAG.

Debe ntụnyere njide ahụ na njedebe njedebe ya. Nsonaazụ gbasara mgbama enwere ike nwetaghachi n'ime oge a nwalere abụghị iwu njedebe zuru ụwa ọnụ. Mkpakọrịta oge nwere ike ịkpali ntụnyere usoro a kapịrị ọnụ na-akọwapụtaghị ihe kpatara ya.

Otu mgbanwe gbanwere Atụrụ anya Gịnị kwesịrị ịnọgide na-adịgide adịgide Oke nke inference
Nhọrọ chromatographic Nkewa na o doro anya na mpaghara ịdị ọcha nwere ike ịbawanye, ibelata, ma ọ bụ nọgide na-adị Ihe mejupụtara ihe atụ, nkwadebe, wee detuo ntọala nchọpụta ebe enwere ike Otu chromatogram dị iche naanị anaghị egosipụta nhazi ogbe dị iche
Ndabere ọrụ ntinye uche Nleba anya agbakọ na nzaghachi nke ahaziri ahazi nwere ike ịgbanwe Ihe n'ezie, olu nkwadebe, na ntụ ntụ Mmetụta ya dabere n'ihe nha ọdịnaya ọ bụla gụnyere
Ogologo oge nkwadebe-iji nwalee Ebumnuche enwere ike nwetaghachi nwere ike ịdịgidere ma ọ bụ gbanwee site na usoro anụ ahụ ma ọ bụ kemịkalụ Akpa, ọkara, okpomọkụ, na usoro nyocha Njikọ oge anaghị achọpụta usoro ọnwụ

Ozugbo mgbanwe dịpụrụ adịpụ, nnwale nke abụọ nwere ike nyochaa usoro ya. Na-agbanwe oge nchekwa, akpa, ihe mgbaze, na itinye uche n'otu ọnụ nwere ike rụpụta nsonaazụ ka mma ma hapụ ihe kpatara amaghị. Nsonaazụ dị otú ahụ nwere ike ịba uru n'ọrụ, mana ọ na-akwado nkwubi okwu dị iche site na ntụnyere otu mgbanwe na-achịkwa.

Kedu ka ekwesịrị isi nyochaa COA na ndekọ anomaly ọnụ?

Gụọ COA dị ka usoro nke nkwupụta nwere njikọ. Ihe nchọpụta ogbe jikọtara akụkọ na vial. Usoro na otu njedebe na-akọwapụta ebumnuche ebumnuche. Ọnọdụ HPLC na-akọwa pasentị mpaghara akọpụtara. Nkọwa uka na-ekwu maka ihe mejupụtara ya, ebe usoro ọdịnaya na-akọwa ọnụọgụ. Nsonaazụ mmiri ma ọ bụ mgbochi na-aza ajụjụ ndị ọzọ naanị ma ọ bụrụ na a tụrụ ya n'ezie.

N'ihi na hypothetical ala mgbama mgbe ijide, buru ụzọ gosi na ejiri otu ogbe na ntuziaka nkwadebe mee ihe. Nyochaa oghere, omume ntụaka, ndekọ dilution, na arụmọrụ ngwá ọrụ. Wee tulee ihe nlele ọhụrụ na nke ejidere na nha nwere ike nyochaa mgbake, kama ịdabere naanị na mpaghara ịdị ọcha nke ọma.

Ọ bụrụ na akwadoro ọnwụ, nyochaa ma enwere ike nwetaghachi ihe n'ime akụkụ ma ọ bụ akpa dị iche yana ma ụdị kemịkal ọhụrụ apụtala. A ga-emepụtarịrị ma kọwagharịa nnwale mgbake maka usoro n'ezie. Ọdịda ịhụ elu ọhụrụ anaghị egosipụta n'onwe ya mgbasa ozi, n'ihi na ụfọdụ ngwaahịa nwere ike ghara achọpụtara nke ọma ma ọ bụ ghara idozi.

Kpalite nyocha ọzọ mgbe njirimara na-ekwekọghị ekwekọ, Ngosipụta adịghị ọcha ekwekọrịtara ada ada, Nleta ịta ahụhụ ekwenyeghị karịa ejighị n'aka ezighi ezi, ma ọ bụ mgbanwe omume omume n'oge mpio ule echere. Kọwaa njirisi dị mkpa na ndị otu ọrụ. Emepụtala ọnụ ụzọ ọdịda ọnụọgụgụ site na ndapụta atụ.

FAQ: na-ahọpụta njikwa maka nyocha peptide sịntetik

Bụ peptide scrambled bụ njikwa mma zuru oke? Mba. Ọ nwere ike na-aza ajụjụ nkọwa nke ndu ma ọ bụrụ na akụrụngwa ya dabara adaba, mana ọ naghị akwado njirimara peptide ule, ọdịnaya, ma ọ bụ ijikwa kwụsie ike. Usoro a gbajiri agbaji anaghị arụ ọrụ ozugbo na usoro ndu.

Oghere nyocha nwere ike gosi na peptide dị ọcha? Mba. Ọ nwere ike ikpughe ndabere, mmetọ, ma ọ bụ ebuga n'okpuru ọnọdụ anwalela. Ọ naghị ewepụta nkewa nke adịghị ọcha metụtara ebumnuche ma ọ bụ njirimara nhazi.

Ịdị ọcha na-agbanweghị agbanwe ọ pụtara itinye uche na-agbanweghị agbanwe? Mba. Ebe ịdị ọcha dị na ya na ego nwere ike nwetaghachi bụ nha dị iche iche. Ihe nlele nwere ike tufuo ihe mgbe ọ na-ejigide pasent mpaghara akọpụtara.

Kedu njikwa kwesịrị ịgbakwunye na mbụ mgbe nsonaazụ ekwenyeghị? Jụọ nke nlebanya ga-amata nkọwa ndị na-eduga. Oghere nwere ike nwale ndabere; ntụaka dabara adaba nwere ike nyochaa akụkụ akọwapụtara nke arụmọrụ nha; ego akaebe na-adabereghị na ya nwere ike ileba anya n'ụkpụrụ ntinye uche. Ọ dịghị onye na-anọchi anya ndị ọzọ zuru ụwa ọnụ.

Zejun Peng

Zejun Peng

Onye isi ọrụ teknụzụ; Ọkachamara Synthesis Peptide Ọkachamara isi: Mgbagwoju anya peptide njikọ, mgbanwe nke amino acid na-abụghị eke, na iwu nke peptides cyclic na peptides stapled.

Akụkọ ndụ:Zejun Peng nwere ahụmahụ dị ukwuu na kemịkalụ organic na njikọ peptide. Ọ maara nke ọma na ngwakọta nke njikọ peptide siri ike-phase (SPSS) na mmiri mmiri-phase peptide njikọ (LPPS), ma bụrụ ọkachamara kachasị n'imeri usoro "usoro siri ike ịmekọrịta" (dị ka peptides ultra-long-chain, nnukwu hydrophobic usoro, na otutu disulfide mpịachi). N'okpuru nduzi ya, otu ahụ nke ọma merie teknuzu bottlenecks na ọtụtụ pụrụ iche mgbanwe (dị ka N-methylation, PEGylation, na akara fluorescent), na-ejigide ọnụ ọgụgụ ihe ịga nke ọma nke ihe karịrị 98%.

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