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Porous silica barbashi da aka shirya ta hanyar sol-gel tare da wasu gyare-gyare don samun macroporous barbashi.These barbashi da aka samu derivatized by reversible Bugu da kari fragmentation sarkar canja wurin (RAFT) polymerization tare da N-phenylmaleimide-methylvinylisocyanate (PMI) da styrene shirya N-phenylmaleimiderrows karfe-polymistyrerrow bakan lokaci (PMI) da kuma styrene don shirya N-phenylmaleimstyrerrow-polymstyrerrow karfe lokaci (PMI). 100 × 1.8 mm id) an cika su ta hanyar slurry packing.Kimanin PMP shafi rabuwa na peptide cakuda kunshi biyar peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin album) da kuma leucine enkephalin. Imal elution yanayi, theoretical farantin kirga na peptide cakuda ne kamar yadda high as 280,000 faranti / m². Kwatanta aikin rabuwa na ci gaba shafi tare da kasuwanci Ascentis Express RP-Amide shafi, an lura da cewa rabuwa yi na PMP shafi ya fi na kasuwanci shafi cikin sharuddan rabuwa dace da ƙuduri.
A cikin 'yan shekarun nan, da biopharmaceutical masana'antu ya zama wani fadada kasuwar duniya tare da wani gagarumin karuwa a kasuwa rabo.Tare da fashewar girma na biopharmaceutical masana'antu1,2,3, da bincike na peptides da sunadaran da ake so sosai. Bugu da kari ga peptide manufa, da dama da ƙazanta ana haifar da a lokacin peptide hadaddun, da yawa najasa da ake samu a lokacin peptide hadaddun, don haka da ake bukata na gina jiki da ake so don samun peptides. ruwa, kyallen takarda da sel wani aiki ne mai wuyar gaske saboda yawancin nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i. spectrometer a lokacin da aka ba da lokaci4,5,6. Bugu da ƙari, a lokacin rabuwar lokaci na ruwa, ana iya mayar da masu bincike a cikin yankuna masu kunkuntar, ta haka ne ke mayar da hankali ga waɗannan nazarin da kuma inganta ƙwarewar ganewar MS. Liquid chromatography (LC) ya ci gaba sosai a cikin shekaru goma da suka wuce kuma ya zama sanannen fasaha a cikin bincike na proteomic7,8,9,10.
Reversed-lokaci ruwa chromatography (RP-LC) ne yadu amfani da tsarkakewa da kuma rabuwa da peptide gaurayawan ta amfani da octadecyl-gyara silica (ODS) a matsayin m Phase11,12,13.Amma, RP tsayayyun bulan ba su samar da m rabuwa na peptides da sunadarai saboda su hadaddun tsarin, 4 amphire tsari da kuma 1 amphire tsari. Ana buƙatar nazarin peptides da sunadarai tare da nau'in polar da marasa iyaka don yin hulɗa tare da kuma riƙe waɗannan analytes16.Mixed-mode chromatography, wanda ke ba da hulɗar multimodal, zai iya zama madadin RP-LC don rabuwa da peptides, sunadaran, da sauran hadaddun hadaddun.Da yawa gauraye-yanayin tsayayye ginshikan da aka yi amfani da wadannan ginshikan da separpt ginshikan da aka yi amfani da furotin 1. 8,19,20,21.Mixed-mode tsayayye matakai (WAX / RPLC, HILIC / RPLC, iyakacin duniya intercalation / RPLC) sun dace da peptide da furotin separations saboda gaban duka iyakacin duniya da kuma wadanda ba iyakacin duniya kungiyoyin22,23,24,25,26,27,28 .Similarly da ikon interparation na musamman polar nuna ƙungiyoyin polar tare da tsaka-tsakin tsaka-tsakin ƙungiyoyin polar na musamman. haɓakawa ga masu nazarin polar da waɗanda ba na polar ba, kamar yadda rabuwa ya dogara da hulɗar tsakanin nazari da lokaci mai tsayi.Multimodal hulɗar 29, 30, 31, 32. Kwanan nan, Zhang et al.30 ya shirya wani dodecyl-terminated polyamine tsayayye lokaci da kuma samu nasarar raba hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, da kuma da dama sauran analytes.The iyakacin duniya intercalator yana da duka iyakacin duniya da kuma wadanda ba iyakacin duniya kungiyoyin, don haka shi za a iya amfani da su raba peptides da sunadaran da cewa suna da duka hydrophobic da kuma hydrophilic-moeties, amiidi kasuwanci Clogin-columns. Akwai a ƙarƙashin sunan kasuwanci Ascentis Express RP-Amide ginshiƙai, amma ana amfani da waɗannan ginshiƙan don nazarin amine 33 kawai.
A cikin binciken na yanzu, an shirya wani lokaci mai tsayi (N-phenylmaleimide-embedded polystyrene) da kuma kimantawa don rabuwa da peptides da trypsin digests na HSA. An shirya lokaci mai tsayi ta hanyar amfani da dabarun da ke gaba. An shirya ƙwayoyin silica da aka yi amfani da su bisa ga tsarin da aka ba a cikin littafinmu na baya tare da wasu gyare-gyaren gyare-gyare na glycol. OS, ruwa acetic acid da aka gyara don shirya silica barbashi tare da babban pore size.Na biyu, wani sabon ligand, phenylmaleimide-methyl vinyl isocyanate, aka hada da kuma amfani da derivatize silica barbashi don shirya wani iyakacin duniya saka a tsaye lokaci.The sakamakon tsayayye lokaci da aka cushe a cikin wani bakin karfe 1.0mm packed shafi (1.0.8). tare da girgizar injiniya don tabbatar da an kafa gado mai kama da juna a cikin ginshiƙi.Yi la'akari da rabe-raben ginshiƙi na cakuda peptide wanda ya ƙunshi peptides biyar;(Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) da kuma Trypsin digest na ɗan adam serum albumin (HAS) .The peptide cakuda da trypsin narkewa na HSA aka lura raba tare da mai kyau ƙuduri da kuma yadda ya dace na PRP da aka kwatanta da ginshiƙin da aka kwatanta da sepa. Dukansu peptides da sunadaran an lura da su don warwarewa da inganci akan ginshiƙin PMP, wanda ya fi dacewa fiye da shafi na Ascentis Express RP-Amide.
PEG (Polyethylene Glycol), Urea, Acetic Acid, Trimethoxy Orthosilicate (TMOS), Trimethoxy Chlorosilane (TMCS), Trypsin, Human Serum Albumin (HSA), Ammonium Chloride, Urea, Hexane Methyldisilazane (HMDS), Methacryloyl Chloride (Mc), Peroxide (Mc), Peroxide, Benzor, Peroxide, Benzor, Peroxide, Benzor, Peroxide (Mc) HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, da Acetone Sayi daga Sigma-Aldrich (St. Louis, MO, Amurka).
A cakuda urea (8 g), polyethylene glycol (8 g), da 8 ml na 0.01 N acetic acid aka zuga na minti 10, sa'an nan 24 ml na TMOS aka kara da shi a karkashin kankara-sanyi yanayi.The dauki cakuda da aka mai tsanani a 40 ° C for 6 hours sa'an nan a 120 ° C da ruwa da aka bushe a cikin ruwa na 8 hours. 0 ° C na tsawon sa'o'i 12. The busassun laushi mai laushi ya kasance ƙasa mai santsi a cikin tanda kuma an sanya shi a 550 ° C na tsawon sa'o'i 12. An shirya batches uku kuma an kwatanta su don nazarin reproducibility a cikin girman girman, girman pore da yanki.
Ta hanyar gyaran gyare-gyare na silica barbashi tare da pre-haɗa ligand phenylmaleimide-methylvinylisocyanate (PCMP) tare da radial polymerization tare da styrene, an shirya wani fili mai kunshe da rukuni na polar.Lokaci na tsaye don tarawa da sarƙoƙi na polystyrene.An kwatanta tsarin shirye-shiryen a ƙasa.
N-phenylmaleimide (200 MG) da methyl vinyl isocyanate (100 MG) an narkar da su a bushe toluene, da kuma 0.1 ml na 2,2′-azoisobutyronitrile (AIBN) aka kara zuwa dauki flask don shirya phenylmaleimide-methyl vinyl isocyanate copolymer. tanda a 40 ° C na 3 hours.
Dried silica barbashi (2 g) aka tarwatsa a bushe toluene (100 ml), zuga da sonicated a cikin wani 500 mL zagaye kasa flask for 10 min.PMCP (10 MG) da aka narkar da a toluene da kuma kara dropwise zuwa ga flask dauki via wani faduwa mazurari.The cakuda aka refluxed a 180 ° C tace da 180 ° C. 3 hours. Sa'an nan, PMCP-bonded silica barbashi (100 g) aka narkar da a toluene (200 ml) da kuma 4-hydroxy-TEMPO (2 mL) da aka kara a gaban 100 µL na dibutyltin dilaurarate a matsayin mai kara kuzari.The cakuda aka zuga a 50 ° C da tace 8 hours.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), da TEMPO-PMCP-haɗe da silica barbashi (1.5 g) an tarwatsa a cikin toluene da kuma tsarkake da nitrogen.The polymerization na styrene da aka gudanar a 100 ° C na 12 hours.Sakamakon samfurin da aka wanke tare da methanol 1 ° C da aka bushe a cikin dare.
An yi amfani da samfurori a 393 K don 1 hour don samun ragowar matsa lamba na kasa da 10-3 Torr. Adadin N2 adsorbed a matsa lamba na P / P0 = 0.99 an yi amfani da shi don ƙayyade jimlar pore. ligand-bonded silica particles) an sanya su a kan wani ginshiƙin aluminum ta amfani da m carbon tef. An yi amfani da zinari a kan samfurori ta amfani da Q150T sputter coater, da kuma 5 nm Au Layer da aka ajiye a kan samfurori.Wannan yana inganta ingantaccen tsari ta amfani da ƙananan ƙarfin lantarki kuma yana samar da hatsi mai kyau, sanyi sputtering.A Thermotham Electron, US Analyly Analyly 1. .A Malvern (Worcestershire, UK) Mastersizer 2000 barbashi size analyzer da aka yi amfani da su samu da barbashi size rarraba.Naked silica barbashi da ligand-bonded silica barbashi (5 MG kowane) aka tarwatsa a cikin 5 mL na isopropanol, sonicated ga 10 min, vortexed ga 5 min, da kuma bene vortexed ga 5 min, da kuma benci. 5 °C a minti daya akan yanayin zafi na 30 zuwa 800 ° C.
Gilashin da aka liyi bakin karfe kunkuntar ginshiƙan girma (100 × 1.8 mm id) an cika su ta amfani da hanyar tattara kayan slurry, ana amfani da wannan hanyar da aka yi amfani da ita a cikin Ref.31.A bakin karfe ginshiƙi (gilashi-layi, 100 × 1.8 mm id) tare da fitting fitting dauke da 1 µm frit da aka haɗa zuwa slurry fakiti (Alltech Deerfield, IL, USA) .Shirya a tsaye lokaci slurry ta dakatar da 150 MG na columnar mL da aka yi amfani da shi a cikin 1.Met. slurry sauran ƙarfi kazalika da propelling solvent.Cika da shafi bi da bi da ake ji matsa lamba na 100 MP for 10 minutes, 80 MP for 15 minutes, and 60 MP for 30 minutes. A lokacin shiryawa, inji vibration da aka yi amfani da biyu GC shafi shakers (Alltech, Deerfield, IL, USA) don tabbatar da marufi a hankali a cikin ginshiƙi marufi Pa. .Cire haɗin ginshiƙi daga naúrar tattara kayan slurry kuma haɗa wani dacewa zuwa shigarwar da kuma tsarin LC don duba aikin sa.
LC famfo (10AD Shimadzu, Japan), injector (Valco (Amurka) C14 W.05) tare da 50nL allura madauki, membrane degasser (Shimadzu DGU-14A), UV-VIS capillary taga da aka gina Special µLC na'urar ganowa (UV-2075) da kuma gilashin-layi layi connected microcolumns da karin gilashi-lined ginshiƙai ƙunci. Bayan marufi, capillaries (50 μm id 365 da kuma rage ƙungiyoyin capillaries (50 μm) aka shigar a 1/16 ″ kanti na rage ƙungiyar. An yi tattara bayanai da sarrafa chromatographic ta amfani da Multichro 2000 software. Kulawa a 254 nm Origin Analytes aka gwada don UVginance data, MA Proampton data aka analyzer.
Albumin daga jinin mutum, lyophilized foda, ≥ 96% (agarose gel electrophoresis) 3 MG gauraye da trypsin (1.5 MG), 4.0 M urea (1 mL), da 0.2 M ammonium bicarbonate (1 mL) . An zuga maganin na minti 10 kuma an ajiye shi a cikin wanka na ruwa a 37 ° C. canza maganin kuma adana a kasa 4 ° C.
An ƙididdige rarrabuwa na cakuda peptide da HSA trypsin digestive daban-daban akan ginshiƙan PMP. Bincika rabuwa da cakuda peptide da trypsin na HSA ta hanyar PMP kuma kwatanta sakamakon zuwa shafi na Ascentis Express RP-Amide. An ƙididdige lambar farantin ka'idar kamar haka:
Hotunan SEM na ɓangarorin silica maras tushe da barbashi na silica masu haɗin ligand ana nuna su a cikin FIG.2. Imagesan hotunan Boye Silica (a, B) sun nuna hakan, sabanin yadda aka ba mu-karawar silica (C, d) ya zama mai narkewa fiye da na ƙwararrun sarƙoƙi na polystyrene.
Ana duba hotunan siliki na siliki (A, B) da ligand-bonded silica (C, D).
The barbashi size rarraba danda silica barbashi da ligand-bonded silica barbashi an nuna a cikin Hoto 3 (A) .Volume tushen barbashi size masu lankwasa ya nuna cewa girman da silica barbashi ya karu bayan sinadaran gyara (Fig. 3A) The barbashi size rarraba bayanai na silica barbashi daga halin yanzu binciken da kuma baya-tushen da aka kwatanta a cikin P. 3.36 μm, idan aka kwatanta da mu baya binciken tare da ad (0.5) darajar 3.05 μm (polystyrene-daure silica barbashi) 34.This tsari yana da narrower barbashi size rarraba idan aka kwatanta da mu baya binciken saboda da sãɓãwar launukansa rabo na PEG, urea, TMOS, da acetic acid a cikin dauki dan kadan girma polystyre barbashi da particle-MP particle. A baya an yi nazari. Wannan yana nufin aikin aikin siliki tare da styrene kawai ya ajiye Layer polystyrene (0.97 µm) akan saman silica, yayin da a cikin lokacin PMP ɗin kauri ya kasance 1.38 µm.
Rarraba girman barbashi (A) da rarraba girman pore (B) na ɓangarorin silica da silica masu ɗaure ligand.
The pore size, pore girma da surface area na silica barbashi na halin yanzu binciken da aka bayar a cikin Table 1 (B) .The PSD profiles na danda silica barbashi da ligand-bonded silica barbashi an nuna a Figure 3 (B) .Sakamako ne m ga mu baya binciken.The pore masu girma dabam na danda da ligand0 nuna 1 part 3 silica, wanda ya nuna 3 part 1 da silic. re size yana raguwa da 69 bayan gyare-gyaren sinadarai, kamar yadda aka nuna a cikin Table 1 (B), kuma an nuna canjin yanayin a cikin Fig. 3 (B) . Hakazalika, ƙananan ƙananan ƙwayoyin silica sun ragu daga 0.67 zuwa 0.58 cm3 / g bayan gyare-gyaren sinadarai. Ƙayyadaddun yanayin yanayin da ake nazarin silica 11 / m2 na baya / m2 (2) wanda aka yi nazari a halin yanzu yana nazarin silica 11 / g2. g) .Kamar yadda aka nuna a cikin Table 1 (B), filin sararin samaniya (m2 / g) na siliki na siliki kuma ya ragu daga 116 m2 / g zuwa 105 m2 / g bayan gyare-gyaren sinadarai.
Sakamakon bincike na farko na lokaci mai tsayi yana nunawa a cikin Table 2. The carbon loading na halin yanzu m lokaci ne 6.35%, wanda shi ne m fiye da carbon loading na mu baya binciken (polystyrene bonded silica barbashi, 7.93% 35 da 10.21%, bi da bi) 42. The carbon loading na halin yanzu m, saboda a halin yanzu postyre loading da SP. ligands irin su phenylmaleimide-methylvinylisocyanate (PCMP) da 4-hydroxy-TEMPO aka yi amfani da.The nitrogen nauyi kashi na halin yanzu m lokaci ne 2.21%, idan aka kwatanta da 0.1735 da 0.85% da nauyin nitrogen a baya karatu, bi da bi.Wannan yana nufin cewa wt % na carbon loading lokaci ne mafi girma a cikin halin yanzu loading lokaci a cikin Siliphem. 4) da (5) sun kasance 2.7% da 2.9%, bi da bi, yayin da carbon loading na karshe samfurin (6) ya kasance 6.35%, kamar yadda aka nuna a cikin Table 2. An duba asarar nauyi tare da PMP tsayayye lokaci, da kuma TGA kwana da aka nuna a cikin Figure 4.TGA curve yana nuna nauyin hasara na 8.6% na carbon, amma 3 ligands ba kawai ya ƙunshi 8.6% ligands ba, amma yana da kyau a cikin 6. da H.
A phenylmaleimide-methylvinylisocyanate ligand aka zaba domin surface gyare-gyare na silica barbashi domin yana da iyakacin duniya phenylmaleimide kungiyoyin da vinylisocynate kungiyoyin.Vinyl isocyanate kungiyoyin iya kara amsa tare da styrene ta rayuwa m polymerization.Dalili na biyu shi ne shigar da wani rukuni da cewa yana da matsakaicin hulda tsakanin analy lokaci da kuma babu wani matsakaicin hulda tsakanin analy tashar da kuma analy lokaci tare da analy tashar. nylmaleimide moiety ba shi da wani kama-da-wane cajin a al'ada pH.The polarity na a tsaye lokaci za a iya sarrafa ta da mafi kyau duka adadin styrene da dauki lokaci na free radical polymerization.The karshe mataki na dauki (free-radical polymerization) ne m kuma zai iya canza polarity na a tsaye phase.Elemental bincike da aka yi don duba da carbon da aka kara loading da adadin lokacin da aka lura da lokacin da aka lura da carbon loading da lokacin da aka lura da lokacin loading. A tsaye lokaci da kuma mataimakin versa.SPs da aka shirya tare da daban-daban taro na styrene da daban-daban carbon loadings.Again, ɗora wadannan a tsaye bulan a cikin bakin karfe ginshikan da duba su chromatographic yi (zaɓi, ƙuduri, N darajar, da dai sauransu) .. Bisa ga waɗannan gwaje-gwajen, an zaɓi ingantaccen tsari don shirya PMP a tsaye lokaci don tabbatar da sarrafawa da polarity da kuma mai kyau analyte.
An kuma kimanta cakuda peptide guda biyar (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) ta amfani da shafi na PMP ta amfani da lokaci na hannu;60/40 (v / v) acetonitrile / ruwa (0.1% TFA) a ma'auni na 80 μL / min. A karkashin yanayi mafi kyau duka, lambar farantin ka'idar (N) a kowane shafi (100 × 1.8 mm id) shine 20,000 ± 100 (200,000 na ginshiƙan chrome / m) yana ba da ginshiƙan 3 na chrome / m. Ana nuna atograms a cikin Hoto 5A. Bincike mai sauri a kan wani shafi na PMP a babban adadin ruwa (700 μL / min), peptides biyar sun ɓace a cikin minti daya, N dabi'u suna da kyau sosai, 13,500 ± 330 a kowace shafi (100 × 1.8 mm id), Daidai da 135,000 ginshiƙai 5 (Siffa 1 × 1) . mm id) an cika su da kuri'a daban-daban guda uku na PMP a tsaye lokaci don duba reproducibility.An rubuta maida hankali na nazari ga kowane shafi ta amfani da mafi kyawun yanayi na elution da adadin faranti na ka'idar N da lokacin riƙewa don raba cakuda gwajin guda ɗaya a kan kowane ginshiƙi. An nuna bayanan reproducibility ga ginshiƙan PMP a cikin Tebura 4. An nuna ma'auni na reproducibility na kowane ginshiƙai tare da ginshiƙan ƙima a cikin ƙimar PMPD sosai.
Rabuwar cakuda peptide akan shafi na PMP (B) da Ascentis Express RP-Amide shafi (A);tsarin wayar hannu 60/40 ACN/H2O (TFA 0.1%), PMP ginshiƙi (100 × 1.8 mm id);Analytical Tsarin elution na mahadi: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) da 5 (leucine) acid enkephalin)).
An kimanta wani shafi na PMP (100 × 1.8 mm id) don rabuwa da gwajin gwaji na jini na jini na ɗan adam a cikin babban aikin ruwa na chromatography. chromatogram a cikin Hoto 6 ya nuna cewa samfurin ya rabu da kyau kuma ƙuduri yana da kyau sosai. An bincikar HSA digests ta hanyar amfani da ƙimar 100 µL / min, motsi na hannu da aka nuna a cikin ruwa na 70.1. chromatogram (Hoto 6), an raba narkewar HSA zuwa kololuwar 17 daidai da peptides 17. An ƙididdige ƙimar rabuwa na kowane kololuwa a cikin HSA narkewa kuma an ba da ƙimar a cikin Table 5.
An raba gwajin gwaji na HSA (100 × 1.8 mm id) akan ginshiƙin PMP;Yawan kwarara (100 µL/min), lokacin wayar hannu 60/40 acetonitrile/ruwa tare da 0.1% TFA.
inda L shine tsayin ginshiƙi, η shine danko na lokaci na wayar hannu, ΔP shine matsi na baya, kuma u shine saurin layi na lokaci na wayar hannu. Ƙaƙƙarfan ra'ayi na PMP ya kasance 2.5 × 10-14 m2, yawan gudu ya kasance 25 μL / min, kuma 60/40 v / v ACN / ruwa ya kasance daidai da 1 × 1.0. na binciken mu na baya Ref.34.The permeability na ginshiƙi cushe da sama da porous barbashi ne: 1.7 × 10-15 ga 1.3 μm barbashi, 3.1 × 10-15 ga 1.7 μm barbashi, 5.2 × 10-15 5.2 × 10-15 da 2.5 μm ga barbashi na 2.5 μm ga 1.5 μm. 43.Saboda haka, ƙaddamarwar lokaci na PMP yana kama da na 5 μm core-shell barbashi.
inda Wx shine nauyin ginshiƙan da aka cika da chloroform, Wy shine nauyin ginshiƙan da aka cika da methanol, kuma ρ shine nauyin ma'auni. Densities of methanol (ρ = 0.7866) da chloroform (ρ = 1.484) .Dukkanin porosity na SILICA PARTICLES-3 × 1 × 1 ginshiƙai 8 × 1 × 1 (0) rea ginshiƙai 31 da muka yi nazari a baya sun kasance 0.63 da 0.55, bi da bi.Wannan yana nufin cewa kasancewar urea ligands yana rage haɓakar lokaci na tsaye. Nau'in tsayayyen matakai C18 ligands suna haɗe zuwa silica barbashi azaman sarƙoƙi na layi, yayin da a cikin nau'ikan nau'ikan polystyrene, an kafa Layer polymer mai kauri a kusa da shi.
Hoto 7A, B yana nuna shafi na PMP (100 × 1.8 mm id) da kuma Ascentis Express RP-Amide shafi (100 × 1.8 mm id) ta amfani da yanayi iri ɗaya (watau 60/40 ACN / H2O da 0.1% TFA).) na makircin van Deemter.Abubuwan cakuda peptide da aka zaɓa (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) an shirya su a cikin 20 µL / Matsakaicin ƙimar kwarara don ginshiƙan biyu shine 800 µL/min. Mafi ƙarancin ƙimar HETPmµ da ƙimar P8 na P shi ne Express RP-Amide shafi sun kasance 2.6 µm da 3.9 µm, bi da bi. Ƙimar HETP suna nuna cewa haɓakar rabuwa na ginshiƙin PMP (100 × 1.8 mm id) ya fi kyau fiye da kasuwancin da ake samuwa Ascentis Express RP-Amide (100 × 1.8 mm id) yana nuna alamar ƙimar da aka nuna a cikin Fig. ba mahimmanci ba idan aka kwatanta da binciken mu na baya. Mafi girman haɓakar rabuwa na PMP shafi (100 × 1.8 mm id) idan aka kwatanta da ginshiƙin Ascentis Express RP-Amide ya dogara ne akan ingantawa a cikin nau'i na nau'i, girman, da kuma hadaddun hanyoyin tattarawa na shafi da aka yi amfani da su a cikin aikin yanzu34.
(A). 0.1% TFA.
A iyakacin duniya-embedded polystyrene m lokaci da aka shirya da kuma kimanta domin rabuwa da roba peptide cakuda da kuma trypsin digests na mutum serum albumin (HAS) a high yi ruwa chromatography.The chromatographic yi na PMP ginshikan ga peptide gaurayawan yana da kyau kwarai a rabuwa yadda ya dace da kuma ƙuduri.The ingantattun rabuwa da ginshiƙai na iya aiki da kuma girman girman girman girman PMP saboda girman girman girman girman PMP. wani barbashi, sarrafawa kira na tsaye lokaci, da kuma hadaddun shafi packing.In Bugu da kari ga high rabuwa yadda ya dace, low shafi baya matsa lamba a high ya kwarara rates ne wani amfani da wannan m Phase.PMP ginshikan nuna mai kyau reproducibility kuma za a iya amfani da bincike na peptide gaurayawan da trypsin narkewa daga daban-daban sunadaran.Mu yi nufin yin amfani da wannan ginshikan na ruwa da kuma bioprotective mahadi daga ginshikan bioprotein. chromatography.A nan gaba, za a kuma kimanta ginshiƙan PMP don rarrabuwar sunadarai da ƙwayoyin rigakafi na monoclonal.
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