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How to store peptides?


Peptides Lyophilized

Mo te rokiroki mo te wa roa o nga peptides, Ka tūtohu mātou kia penapena hei paura mārō. Ka taea te penapena i nga peptides Lyophilized ki te -20 ° C, ki raro ranei me te iti o te whakaheke. Ka tohutohuhia nga kaihoko ki te tirotiro ano i nga peptides kua whakaritea ia 2 tau mai i te ra o te tukunga mana kounga.

Peptides i roto i te otinga

Ko nga peptides i roto i te otinga he iti ake te pumau, he ngawari ki te paheketanga. Ko nga wairewa wai e whakamahia ana ki te whakakore i nga peptides me parakore me te parakore.

I roto i te otinga, ka taea te whakaheke i tetahi nui o te peptide, i runga i nga waikawa amino kei roto i te raupapa. Ko etahi tauira kei raro nei:
– Peptides kei roto te methionine, Ko nga toenga o te cysteine, tryptophan ranei, me iti te wa rokiroki i roto i te otinga na te mea te waikura. Me rewa enei peptides i roto i te whakarewa hāora-kore. –
Ka taea te whakakore i te Glutamine me te asparagine ki te waikawa glutamic me te waikawa aspartic, ia. –
Ka taea e Cysteine ​​​​te paheketanga oxidative ki te hanga i te here cysteine-cysteine ​​disulfide (ka taea te hanga i roto, i waenganui ranei i nga here disulfide). –
Ko nga toenga kua tohua (waikawa aspartic, waikawa glutamic, raihine, arginine, histidine) he hygroscopic (te tango wai mai i te hau) me te kaha ki te hanga viscous, hinu marama. Kaore pea tenei huringa tinana e pa ki te ahua o te peptide. E taunaki ana kia lyophilized aua peptides hei tango i te nui o te wai, katahi ka whakahekehia ki te hauota kia kore ai he kohu wai kei roto i te ipu, katahi ka tere te kati.

Hei aukati i te kino mai i te whakatoo me te rewa, Ka tūtohu matou kia whakakorehia e nga kaiwhakamahi te nui o te peptide e hiahiatia ana mo te whakamatautau o naianei. Me penapena te taikaha o te otinga ki te ≥ -20 o C tae noa ki te hiahiatia.

A ape i te makuku.
I te mea ka tino whakaitihia e te makuku te pumau o nga peptides mo te wa roa, Ka tūtohu mātou kia whakaōrite i ngā peptides i roto i te desiccator ki te pāmahana rūma i mua i te whakatuwheratanga o nga ipu. I te otinga o te tuku peptide, Ko nga toenga peptides i roto i te ngongo me ata horoi ki te hauota hauota, katahi ka kapi te ipu, hiritia ki te kiriata hiri, ka rongoa ki te -20°C.


He rereke nga ahuatanga o te whakarewatanga o nga peptides rereke. Ko nga toenga penei i te alanine (Ala), hauteine (Cys), isoleucine (Me), riihi (Leu), methionine (I tutaki), phenylalanine (Phe), me valine (Val) tāpiri ki te hauwai o te peptide me te wairewa i roto i te wairewa wai, a ka tūtohuhia kia whai te kaihoko i enei aratohu i te wa e whakarite ana i nga peptides.

Ngā Āhuatanga Wairewa

Ko te whakarewatanga o nga peptides e tino whakawhirinaki ana ki te raupapa waikawa amino. Nga peptides hydrophobic taiao (A, F, G, V, L, I, M, W, P me te kaha ake) e hiahia ana nga whakarewa pararopi mo te whakarewatanga. Peptides waikawa (teitei ake mo D, E i roto i te raupapa peptide) e hiahia ana ki nga papaa wai mo te whakarewatanga, i te peptides taketake (teitei ake mo te K, H, me R) e hiahia ana ki te wai whakaerenga waikawa mo te whakarewatanga.

Te whiriwhiri whakarewa

I runga i nga here o to whakamatautau, Ka tūtohu matou ki te whakamahi i nga aratohu e whai ake nei hei whakatau i te whakarewa pai mo te whakarewa i nga peptides

Peptides Hydrophobic

Hei whakarewa ano i nga peptides hydrophobic, tāpiri 100 µL o DMSO me te sonicate tae noa ki te hanga i tetahi otinga rite. Whai muri, tāpirihia te parepare e hiahia ana koe ki te hanga a 1 mg/mL otinga (te teitei o te kukū peptide, ko te nui ake o te DMSO e hiahiatia ana).

Waiwai (Waikawa) Peptides

Hei whakarewa ano i nga peptides waikawa, tāpiri 100 µL o 1% NH 4 OH ki 1 mg o te peptide me te vortex. Kia marama te otinga, tāpirihia te parepare e hiahia ana koe ki te hanga a 1 mg/mL otinga.

Waiwai (Taketake) Peptides

Hei hanga i te peptide taketake, tāpirihia te wai māturu ki 1 mg o te peptide me te vortex.


Ko te paerewa ahumahi ko te tuku peptides i roto i te ahua lyophilized hei taumaha Gross. Te taumaha peptide nui Ko te taumaha katoa o nga waahanga katoa kei roto i te paura lyophilized. Kei roto i tenei ko te peptide o te paanga, tetahi poke peptide, wai, whakarewa toenga, me te taupatupatu.

Engari Kupenga taimaha peptide, tatauhia mai i te Te ihirangi peptide kupenga, ko te taumaha o ko te peptide anake he waahanga kei roto i te paura lyophilized. Ko tenei ka nui ake te tika o nga kohinga peptide o muri mai. No reira, I taunakihia e matou nga moni taumaha peptide kupenga mo Peptide Ritenga whakahau.

Net Peptide Content Whakaahua

Ihirangi Peptide Net Ka mahia te ine ma te tātari waikawa amino (AAA; iti te tika engari me iti te rahinga rauemi) te tātari huānga rānei (CHN; ka hiahiatia he milligrams o te peptide engari he tika ake). Ko nga tikanga e rua ka puta he uara ōrau (arā., 75% ihirangi peptide kupenga).

Ko te Ihirangi Peptide Net e whakawhirinaki ana ki te nui o te pareparenga kei reira*, e whakawhirinaki ana ki te raupapa peptide. Ko te tikanga, teitei ake te utu peptide, ka iti ake te ihirangi peptide

*Tuhipoka: Ko te huānga e whakamahia ana ki te tatau i te ihirangi peptide kupenga kei runga i te peptide counter-ion. Ka mahia tenei ki te whakarite kia kore te katote-a-tete e uru ki te nui o te ihirangi peptide. Hei tauira, i te wa ko TFA, acetate ranei te katote-atete, ka whakamahia te hauota na te mea ka kitea i roto i te peptide engari kaore i roto i te TFA, i te acetate ranei. Ka taea te whakamahi i te waro, te Nitrogen ranei i te mea ko te hauota te katote-atete mo taua take ano.

Tataunga Taumaha Peptide Net:

Te Rahi Peptide Kupenga (peptide katoa) - te tikanga e whakamahia ana e matou mo nga ota peptide ritenga kupenga.

Whakareatia te Ōrau Ihirangi Peptide Net (puka ā-ira) na te Peptide Gross Weight, ki te whiwhi i te nui o te Rahi Peptide Net (peptide katoa).

Hei tauira:
Mo 5 mg peptide / ipu:
Ko te ihirangi kupenga Peptide 84%.
Te rahinga o te tapeke peptide = 0.84*5 mg=4.2 mg te peptide katoa

Peptide Net of Interest Rahi—tino tika - Ka taea e te kaihoko te whakamahi i tenei tikanga ki a raatau ake ki te tatau i te peptide o te paanga.

Whakareatia te ōrau Ihirangi Peptide Net ki te Taumaha Kere Peptide, me te Pure Peptide (na HPLC), ki te tatau i te rahinga tika o te peptide o te paanga kei a koe. Te ōrau peptide kupenga (puka ā-ira) ka whakanuia e te pure peptide (puka ā-ira).

Hei tauira:
Mo 5 mg peptide / ipu kerekere ki:
Ko te ihirangi kupenga Peptide 84%.
Ko te nui o te peptide o te huamoni ko = 0.84*0.96*5 mg=4.03 mg


Ka whakamahia e nga Huringa MOL te matū Fmoc-waa totoka, kaua te matū BOC. Ka taea e te matū Fmoc te tango i nga roopu tiaki ma te whakamahi i nga tikanga ngawari i te wa e roa ana te peptide, i te wa e hiahia ana te matū BOC ki te whakamahi i te waikawa kaha ki te tango i nga roopu tiaki i te wa e roa ana te peptide. I tua atu, Ka whakamahia e te matū Fmoc te waikawa trifluoroacetic (TFA) ki te tango i te peptide mai i te kapia, i te wa e whakamahia ana e te matū BOC te waikawa hydrofluoric (HF). He taekore te HF, tino pirau, me te paihana whakapiri.

No reira, Ka whakamahia e MOL Changes te matū Fmoc na te mea he ngawari ake, ngawari, me te maha o nga mea ka whakaritea ki te matū BOC.


Molarity e pā ana ki te kukū molar o te otinga, ko te maha o nga kiore o te wairewa kua memeha ki roto 1 rita o te otinga, kua whakahuahia hei mol/L, ko M.

Molarity [M] = Mass / (Volume x Molar Mass); Kamiriona = Concentration (g/L) x Rōrahi (L)/MW (g/mol)

Tauira:
Ka hoatu: 1mg o te paura peptide maroke me te MW: 20KDa (Molar papatipu o peptide he 20 g/mol)
Hei whakatau i te molarity me te papatipu mohio me te rōrahi e mohiotia ana
Mo te otinga 1ml o tenei peptide:
Molarity = Mass (0.001g) / (rōrahi (0.001L) x Molar Mass (MW 20,000) = 50 µM

Ki te whakatau papatipu ki te whakatutuki i tetahi molarity:
Mena mo to whakamatautau, me 0.01mM te otinga peptide mahi i roto i te 1ml wai, ka tatauhia te papatipu e hiahiatia ana penei:

Papatipu = Molarity x Volume x Molar Mass
Papatipu = 0.01mM x (1/1000 L) x 20,000g/mol = 0.0002g

No reira, ka hiahia koe 0.2mg/ml o peptide ki te whai i te otinga mahi o 0.01mM.


Ka tukuna e matou hei lyophilized, raw paura ranei me te otinga peptide.


Ko te memehatanga o te peptide i roto i tetahi whakarewa ka whakawhirinaki ki te raupapa waikawa amino, he uaua ki te matapae.. I roto i te take o peptides putumōhio, the solubility information is listed on the QC datasheet. For custom peptides, please see the section entitled “How should I solubilize my peptide”.


Peptide content is not an indication of the peptide purity as these are two separate measurements. Purity is determined by analytical HPLC and indicates the presence/absence of other peptidic contaminants (i.e. truncated peptides).

Net peptide content determines the actual amount of the peptide (peptide of interest as well as the peptide contaminants) present in the sample. The net peptide content is traditionally measured by Amino acid analysis (AAA; limited accuracy but requires a low amount of material) te tātari huānga rānei (CHN; ka hiahiatia he milligrams o te peptide engari he tika ake).


MOL Huringa peptides te tikanga pure e te whakamuri-wāhanga takarepa HPLC, te whakamahi 2 pūreirei kei roto 0.1% waikawa trifluoroacetic (TFA). Ko te parepare tuatahi (Kaipupuri A) he mea tito 0.1 % TFA i roto i te wai whakaheke me te parepare tuarua (Kaipupuri B) he mea tito 0.1 % TFA i roto i te acetonitrile (ACN).

Ko te tikanga, Ko nga peptides masivesivá ka rewa tuatahi ki te otinga o te Buffer A, he nui o Buffer B, he whakarewa polar waro ranei penei i te DMSO. Mena i whakamahia he whakarewa polar organic, Buffer B ranei, Ko tenei otinga marama ka waimeha tuatahi ki te Buffer A. Whai muri, ka tātarihia te otinga peptidic ka utaina ki runga i te punaha HPLC whakarite. Te whakamahi i te rōnaki tikanga arotau, ka kohia nga hautau i runga i nga panui whakauru. Ka tātarihia ia hautau ma te HPLC tātari hei whakarite kia tutuki te ma o ia hautau ki te whakaritenga parakore e hiahiatia ana..

Ka whakahiatohia enei hautau "parakore" ka lyophilized. Ko te paura "parakore" ka whakamatauhia mo te ma me te tuakiri kia mohio ai kua hangaia te peptide tika..

Ka mutu, he roopu QC motuhake i AnaSpec ka whakamatau ano i te peptide hei whakarite kia tutuki katoa nga huanga whakamatautau ki nga whakaritenga e hiahiatia ana.


Ko te whakaingoatanga o te ao e whakamahia ana mo te waahanga raupapa e whai ake nei:

– N-mutu: H te tikanga amine kore utu (NH2-), Ac tikanga acetyl [CH3C(O)-NH-], Pyr tikanga pyroglutamic acid
– C-terminal: Ko te tikanga o te OH he waikawa kore utu (-COOH), Ko te NH2 te tikanga o te amide [-CONH2]

– Ko nga whakarereketanga i te taha taha o nga waikawa amino e whakaatuhia ana i te reu i muri i te waikawa amino e rite ana.. Hei tauira; serine phosphorylated = S(PO3H2) ko te epsilon-N-acetylated lysine = K(Ac)


I te wa o te hanga peptide, Ka herea nga katote i roto i te whakahiato me te purenga ki nga mekameka taha waikawa amino o te peptide., hanga tote. Ko enei katote e mohiotia ana ko nga peptide counterions ka awhina i te whakataurite i te utu o te peptide.

Te waikawa trifluoroacetic (TFA) Kei te whakamahia nuitia i roto i te peptide synthesis me te purenga.

Ka hangaia he tote trifluoroacetate me nga waikawa amino taketake (e.g., raihine, arginine, histidine) me te amine N-terminal kore utu. No reira, trifluoroacetate ko te tikanga te parepare taunoa.

Ko te iti rawa o nga ngota ngota TFA e herea ana ki tetahi peptide he rite ki te maha o nga toenga taketake i roto i te raupapa peptide kua tohua.. Koia, ko nga toenga taketake kei roto i te peptide, ka iti ake te ihirangi peptide kupenga o te paura lyophilized. Ki te whakaaro mo te TFA me te ihirangi wai kei roto i te tauira, Ka tūtohu matou ki te mahi i tetahi whakamatautau ihirangi peptide mo ia peptide ritenga.

Te wai me te waikawa trifluoroacetic (TFA) he hua o te tukanga purenga. Heoi ano, Ka ara ake pea a TFA i nga awangawanga paitini mo ora me nga rangahau preclinical. No reira, Ko nga Huringa MOL e tuku acetate kore-paitini, pūhaumāota, he whakawhiti tote haukinia ranei.


Peptides kei roto Cys, I tutaki, kei te wero ranei a Trp ki te whakahiato me te whiwhi hua tino parakore. Ko te nuinga o tenei na te koretake me te whakaraerae ki te waikura o enei roopu mahi. Me aro nui ki te whakamahi me te rokiroki o enei peptides ki te karo i te whakatuwheratanga o nga ipu.

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hoki mai ki roto 24 haora.

 

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