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Mmepụta mgbochi ọrịa sitere na peptides sịntetik na-achọ imepụta reagents megide peptide antigen akọwapụtara, mana nnabata nke peptide ahụ anaghị akpaghị aka…
Mmepụta mgbochi ọrịa na Peptides sịntetik - Njikwa maka nnabata Peptide na Protein
Mmepụta mgbochi ọrịa sitere na peptides sịntetik na-achọ imepụta reagents megide peptide antigen akọwapụtara, mana nnabata nke peptide ahụ anaghị ewepụta nrịbama nke protein isi n'ime nyocha a chọrọ. Protein nwere ike igosi usoro dị iche iche, mgbe ebu, njikọ, ma ọ bụ akụkụ ndị ọzọ nke immunogen nwere ike inyekwu nzaghachi mgbochi mmadụ. Ya mere atụmatụ bara uru na-ekewapụta ihe owuwu ọgwụ mgbochi, nsonaazụ nyocha peptide, na ihe akaebe nke ngwa-ngwa maka ezigbo protein ebumnuche.
Atumatu ntule: otu otu na-ahọrọ akụkụ ahịrị site na protein nyocha iji mepụta ihe mgbochi maka immunoblotting. Atụmatụ ọrụ gụnyere njikọ na njikọ ụgbọelu. Ndị otu a nwekwara olile anya iji reagent emechaa maka ijide protein-protein, ọ bụ ezie na enyochabeghị ngwa nke abụọ ahụ.
Enweghị nzaghachi mgbochi mmadụ, nkọwapụta, ma ọ bụ achọpụtara nke ọma. Nyochaa imewe kwesịrị igosipụta nke antigen na njikwa dị mkpa, wee kọwaa ihe akaebe ga-akwado ngwa ọ bụla ezubere. Nsonaazụ peptide dị elu naanị enweghị ike ịza ajụjụ ndị ahụ niile.
Jiri tebụl okwu mee ka ọkwa akaebe dị iche iche
| Oge | Pụtara na ọrụ a | Ihe ọ na-adịghị egosi naanị ya |
|---|---|---|
| Peptide antigen | Usoro akọwara na ụdị molekụla ahọpụtara dị ka mpaghara nnabata ndoro-ndoro ochichi | Na mpaghara ahụ nwere ike ịnweta ma ọ bụ gosi otu a na nyocha protein |
| Immunogen nke na-ebubu | Agbakwunyere peptide site na kemistri akọwapụtara na onye na-ebu ya | Na nzaghachi ọ bụla na-apụta bụ naanị na mpaghara peptide ezubere |
| Aka njikọ | Otu ìgwè eji ejikọta peptide, nke nwere ike ịbụ nwa afọ ma ọ bụ ụma tinye ya | Na ọ bụ nke usoro protein izizi |
| Ihuenyo na-ekekọta peptide | Ntụle njirimara n'ụdị nyocha akọwapụtara | Nkwenye kpọmkwem maka protein isi iyi na ngwa ọ bụla |
| Ngwa-kpọmkwem nkwado | Ihe akaebe na-ekwu maka arụmọrụ reagent n'ime nyocha echere na ọnọdụ nlele | Ịrụ ọrụ zuru ụwa ọnụ na usoro ndị ọzọ, ihe atụ, ma ọ bụ ọtụtụ |
| Mgbochi ma ọ bụ nlele asọmpi | Nsonaazụ na-egosi ka antigen agbakwunyere si emetụta mgbama atụpụtara | Mwepu zuru oke nke mmegharị mmegharị ya na epitopes ndị ọzọ metụtara ya |
Ihe dị iche iche dị mkpa mgbe otu ọrụ si na immunoblot gaa na nyocha nke nwa afọ-protein. Ọnọdụ anụ ahụ na nnweta nke ebumnuche nwere ike ịdị iche. Ihe mgbochi nwere ike ịba uru n'otu ọnọdụ na-egosighi uru na nke ọzọ.
Uhlen na ndị ọrụ ibe nkwado nkwado-mmadụ na-emesi ihe akaebe ike n'ime ngwa a chọrọ na ọnọdụ. Ọ na-akwado ịhazi ntụnyere metụtara ebumnuche ikpeazụ, kama ịgwọ ihuenyo peptide dị elu dị ka nkwado zuru oke maka ma immunoblotting na njide ala.
Ezipụta peptide na conjugation na-agbanweghị epitope e bu n'obi nọrọ jụụ
Nye ihe nchọpụta protein isi iyi na ụdị, nọmba fọduru, kpọmkwem usoro ahọpụtara, ọnụ ọnụ, na mgbanwe ọ bụla ezubere. Ọ bụrụ na ọrụ a metụtara otu ụdị ahaziri ma ọ bụ ọnọdụ mgbanwe, kọwaa ya nke ọma. Enwere ike ịkọwa otu mpaghara mkpụmkpụ ahụ n'ụzọ dị iche mgbe ihe ntụnyere aka ya na-efu.
Hexapeptide 1 Nhazi ihe atụ. Ọ bụrụ na akụkụ ahọpụtara na-ekpuchi ihe fọdụrụ nne na nna 51-64 yana mgbakwunye mgbakwunye cysteine na njedebe N ya., ihe fọdụrụ nke mbụ arụpụtara abụghị nne na nna Cys50. Maapụ ihe fọdụrụ nke mbụ nye nne na nna 51 ọbụna ma ọ bụrụ na atụmatụ ọnụọgụ antigen mpaghara na-atụgharị ya na ọnọdụ 2. Chekwaa maapụ abụọ a ka a na-akọ na njirimara peptide agaghị abụ nkwupụta ụgha gbasara usoro protein isi mmalite.
Jiri nrịbama ụdị ebumnuche doro anya yana maapụ antigen enwere ike ịgụ. Chọpụta akụkụ protein nke ala na cysteine ọ bụla agbakwunyere, spacer ma ọ bụ okpu eji eme njikọ. Chekwaa njikọ ọnụọgụ na mgbakwunye injinia n'ime faịlụ nhazi kama idetuo mgbakwunye ndị ahụ na usoro protein nne na nna..
Ụlọ ọrụ Thermo Fisher amine-reactive chemistry ndu na-achọpụta N-terminal na lysine primary amines. Ọtụtụ amines dị nwere ike ịmepụta ajụjụ saịtị mgbakwunye. "Amine jikọtara" na-akọwa otu klas kemistri; ọ naghị akọwapụta otu ngosi mgbe ọtụtụ saịtị nwere ike meghachi omume.
Jụọ kemịkalụ mgbakwunye na nkesa saịtị echere, otú e ji mara conjugate, na nke ejighị n'aka na-anọgide. Nsonaazụ njirimara peptide na ntinye ma ọ bụ ajụjụ nkesa conjugate bụ nha dị iche iche. Echela na otu uka peptide n'efu na-akọwapụta conjugate nke ụgbọelu niile.
Nke ibe nkwadebe antigen nke Monash na-ekwu na peptide njikọ, Enwere ike ịhazi nchacha na njikọta site n'aka onye na-eweta ngwaahịa masịrị. Nke a na-egosi ụzọ ọrụ ahaziri ahazi, Ọ bụghị nkwa nzaghachi mmadụ ma ọ bụ nkwupụta na ikpo okwu na-emepụta peptides n'onwe ya.
Ihuenyo recombinant-protein nwekwara ike iwebata ihe ọzọ metụtara mkpado. Otu ibe Monash na-akọwa ntụnyere na protein na-adịghị mkpa nwere otu mkpado. Ọ bụrụ na onye ndoro-ndoro na-amata protein abụọ nwere mkpado, enweghị ike ịkenye nsonaazụ na ebumnuche a chọrọ ka ọ dị iche na usoro ala ya. Ebumnuche na-enweghị mkpado dabara adaba ma ọ bụ ihe ngosi ọzọ ziri ezi nwere ike ịza ajụjụ dị iche, mgbe ị na-ejigide njikwa ndị achọrọ maka nyocha n'ezie.
Peptide vs Amino Acid Nọgidenụ na-enyocha reagents n'ọkwa ahụ: ụdị protein, mkpado fusion, ebu, njikọ na otutu. Ịkpọ nke ọ bụla nanị antigen na-ehichapụ ihe dị iche iche na-eme ka njikwa ndị ahụ nwee ike ịkọwa. Nnyefe ahụ kwesịrị ikwe ka ụlọ nyocha ọzọ kọwaa njirimara ekesara na nke agbanwere na ntụnyere ọ bụla.
Ogologo Peptide, solubility, na a ga-enyocha ike nchikota maka onye ndoro-ndoro ochichi. Ọ nweghị ogologo zuru ụwa ọnụ ma ọ bụ pasent ịdị ọcha na-ekwe nkwa ọgwụ mgbochi bara uru. Amụma usoro-nhọrọ bụ ntinye imewe, ọ bụghị ihe ngosipụta gosipụtara na nyocha ikpeazụ.
Wụpụta njikwa na-elebara peptide anya, ebu, na protein ajụjụ
| Ngwa mkpa | Nkọwa nkọwa | Njikwa igodo ma ọ bụ comparators | Nkọwa nke ọdịda |
|---|---|---|---|
| Nyochaa njirimara peptide | Akọwapụtara peptide efu na ihe ngosi nyocha akọwapụtara nke ọma | Nzụlite kwesịrị ekwesị na ntụnyere peptide na-enweghị njikọ | Nsonaazụ adịghị ike nwere ike ịgụnye ngosipụta peptide ma ọ bụ nzaghachi reagent, ọ bụchaghị protein isi iyi |
| Nyochaa mgbama metụtara ụgbọ | njirimara ebu, njikọ kemistri, na mkpa conjugate ozi | Njikwa ihe metụtara ụgbọelu na njikwa ihe ngosi dabara na ihuenyo | Enweghị ike ịkenye mgbaama nye ndị na-ebu ibu na-ekerịta iche na peptide ezubere iche |
| Chọpụta ebumnuche site na immunoblot | Onye ndoro-ndoro ochichi nwere ụdị protein e bu n'obi na ọnọdụ nlele anya | Atụmatụ nkwado dabara adaba na njikwa nyocha dabara adaba | Njikọ Peptide anaghị ekwe nkwa mgbama protein a na-atụ anya ya |
| Weghara protein nke ala | Kewapụ nke ọma ngwa echere na ajụjụ antigen-iche dị mkpa | Njikwa na-ekwu maka mkpakọrịta ndị ebumnuche obodo na ndabere | Nsonaazụ immunoblot enweghị ike igosipụta arụmọrụ njide nke ala |
| Tulee ọtụtụ reagents | Ihe mgbochi a na-achọpụta, peptide, na conjugate ihe nchọpụta ebe ọ dị mkpa | Ntụaka nkịtị na otu ngwa akọwapụtara | A ga-ekewapụ ọtụtụ ihe dị iche iche na mgbanwe nlele na nyocha |
Nleba anya nke na-eji otu ihe na-ebu ya n'ọnọdụ ọ bụla nwere ike ịhapụ nzaghachi metụtara ụgbọelu siri ike ịmata. Ngosipụta dị iche ma ọ bụ njikwa ebu kwesịrị ekwesị nwere ike zaa nchegbu ahụ, ma a ga-enyocha nhọrọ ahụ maka nyocha ahụ n'ezie karịa ka a na-ewere ya dị ka ihe ngbanwe zuru ụwa ọnụ.
Jụọ ihe ntụnyere ụgbọelu-mgbanwe na-ewepụ n'ezie
Tụlee ihuenyo echiche site na iji peptide P jikọtara na onye na-ebu A maka ọgwụ mgbochi yana otu peptide ahụ na-ebu B maka nyocha.. Nkwenye nke P-B na nzaghachi dị ala na B naanị nwere ike ibelata nchegbu Ọdịdị nke Peptide gbasara nnabata a na-eduzi naanị onye na-ebu B. Nnwale A naanị na-ekwu maka usoro ọzọ metụtara ụgbọelu. Nnwale pụrụ iche na nzụlite ka chọrọ nlebara anya maka ntụnyere ahụ.
Ọbụlagodi njikwa ụgbọ dị mma anaghị ewepu atụmatụ ọ bụla na-ekekọrịta. Ọ bụrụ na njikọ abụọ ahụ na-eji otu njikọ ahụ ma ọ bụ mepụta ihe ejikọtara na-ejikọta ya, reagent nwere ike ịmata njirimara ahụ ma ọ bụ ihe ngosi peptide-plus-linker. Ngosipụta ọzọ ziri ezi nwere ike ileba anya n'echiche ahụ. Akọwakwala mgba ọkụ ụgbọelu ka ọ na-ewepụ nsogbu ntụgharị okwu niile metụtara njikọ.
Nchọpụta peptide efu na-enye nleba anya dị iche, ma ka na-adịghị guzobe nnabata nke protein isi iyi adịghị. Mpaghara enwere ike ịnweta na peptide dị mkpụmkpụ nwere ike ịbụ Hexapeptide 10 ewepụtara dị iche iche na protein apịaji ma ọ bụ dị naanị mgbe denaturation gasịrị. Debe nsonaazụ ọ bụla dị mma site na ihe na ngwa a tụrụ ya.
N'otu aka ahụ, peptide enweghị njikọ bara uru naanị maka ajụjụ ọ na-aza. Ọ nwere ike inye aka ikpughe njikọ sara mbara na-enweghị nkọwa na-ewepughị nnabata nke usoro nwere njikọ chiri anya na protein ọzọ.. Kọwaa ebumnuche ọzọ dị mkpa mgbe nkọwapụta dị n'etiti ngwa ahụ.
Debe ọrụ nchịkwa dị mma iche na immunogen. Njikwa na-egosi na usoro nyocha ahụ na-arụ ọrụ anaghị egosi na onye na-eme ihe mgbochi na-amata protein a chọrọ. Mkpesa ikpeazụ chọrọ ihe akaebe jikọtara na mmekọrịta ebumnuche ahụ.
Nyochaa reagenti ikpeazụ n'ọnọdụ nlele echere
Maka oru ngo immunoblot hypothetical, họrọ atụmatụ nkwado dabara adaba na ndị otu nyocha. Uhlen na ndị ọrụ ibe na-ekwurịta ụzọ dị ka mkpụrụ ndụ ihe nketa na ihe akaebe nke orthogonal. Jiri atụmatụ dabara adaba na ebumnuche na ihe gbara ya gburugburu kama ịdegharị aha usoro n'ime akụkọ na-enweghị ntụnyere nkwado. Hexapeptide 11
Nnwale mgbochi nwere ike ịkwado nkọwa metụtara antigen, mana ọ nweghị ike iwepụ naanị mmeghari mmegharị ya na epitopes yiri ya. N'otu aka ahụ, otu band na nha anya a na-atụ anya bụ ihe nlele iji nyochaa, ọbụghị ihe akaebe zuru oke nke njirimara ebumnuche.
Asọmpi nwere ike wepu mgbaama na nnabata nke ebumnobi ebu n'obi kesara yana molekul ọzọ na-ebu epitope yiri ya. Ya mere, eriri egbochiri dakọtara na mmekọrịta metụtara antigen mana ọ naghị achọpụta molekul ọ bụla na-enye aka na band ahụ.. Atụmatụ nkwado metụtara ebumnuche, dị ka ntụnyere mkpụrụ ndụ ihe nketa ma ọ bụ nke nwere onwe ya kwesịrị ekwesị, na-akọwa akụkụ dị iche iche nke ajụjụ nkọwapụta n'okpuru ngwa akọwapụtara. Lipopeptides
Ọ bụrụ na njide nwa amaala ghọrọ ngwa achọrọ, kọwapụta ma nyochaa ya iche. Ihe ndekọ peptide mbụ na ndekọ conjugate ka bara uru, mana ha anaghị edochi anya maka ihe akaebe sitere na ihe gbasara ala-protein. Dekọọ reagent immunoblot bara uru dị ka nke a ruo mgbe akwadoro ngwa ọzọ.
Chekwaa nkọwapụta antigen, ozi njikọ, peptide na antibody otutu identifiers, ọnọdụ nyocha, data nyocha mbụ, njikwa, na mkpebi nyocha. Ụlọ nyocha na-anata kwesịrị inwe ike ịchọpụta ihe reagent n'ezie yana ọnọdụ ebe a tụlere arụmọrụ ya.
Kweekọrịta na ngwa mbụ, ihe ngosi antigen, Nhọrọ ndị dị mkpa na ihe akaebe ikpeazụ metụtara ebumnuche tupu ịnye ọrụ ahụ. Were njide nwa afọ dịka ihe ọzọ mgbe ọ dị mkpa. Dekọọ arụmọrụ na-aga nke ọma n'ogo egosiri kama ịgbasa otu nsonaazụ peptide na-ekekọta n'ime nnabata protein zuru ụwa ọnụ.
Ajụjụ a na-ajụkarị
Na-eme njikọ siri ike na peptide sịntetik na-ekwe nkwa nnabata protein? Mba. Isi iyi protein na nyocha nwere ike igosi mpaghara ahụ n'ụzọ dị iche. Nyochaa ihe mgbaru ọsọ n'ezie na ngwa e zubere na ọnọdụ nlele.
Ekwesịrị ka akọwara cysteine njikọ agbakwunyere dị ka akụkụ nke usoro protein? Mba. Kanye ya dị ka ihe mgbakwunye agbakwunyere ma chekwaa ihe dị iche na ihe fọdụrụ n'ime obodo. Ọrụ ya na ọnọdụ ya kwesịrị ịpụta ìhè na nkọwa antigen.
Onye na-ebu ụgbọ nwere ike ịmepụta nkọwapụta mgbochi mmadụ zuru oke? Mba. Ọ na-agwa mgbama metụtara ụgbọelu. Ajụjụ ndị ọzọ dị mkpa na-apụ apụ ma ọ bụ ihe ngosi chọrọ akaebe nke ha.
Immunblotting na-aga nke ọma na-akwado ijide protein-protein nke ala? Ọ bụghị na-akpaghị aka. Mesoo ngwa nke abụọ dị ka nkwupụta arụmọrụ dị iche site na iji njikwa dabara adaba na data nkwado.
Tupu ịtụ, kwado ajụjụ atọ ndị a:
- Kedu ụdị protein na nyocha nke ọgwụ mgbochi ga-amata?
- Kedu ka a ga-esi tinye peptide ahụ, na nke njikwa iche peptide, ebu, na nzaghachi ngosi?
- Kedu ihe akaebe akọwapụtara nke ngwa ga-akwado nnabata nke reagent mgbochi ikpeazụ?
