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Silica barbashi da aka shirya ta hanyar sol-gel tare da wasu gyare-gyare don samun macroporous barbashi.Waɗannan barbashi da aka samu derivatized by reversible Bugu da kari fragmentation sarkar canja wuri (RAFT) polymerization tare da N-phenylmaleimide-methylvinylisocyanate (PMI) da styrene don shirya N-phenylmaleimstyrerrow-polymiderrow lokaci (PMI). ginshiƙan bakin karfe (100 × 1.8 mm id) an cika su ta hanyar slurry packing.Kimanin PMP shafi rabuwa na peptide cakuda da ya ƙunshi peptides biyar (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Argographic na mutum-mutum, gwada gwadawa). serum albumin (HAS) .A karkashin mafi kyau duka yanayi elution, theoretical farantin count na peptide cakuda ne kamar yadda high as 280,000 faranti / m². Kwatanta da rabuwa yi na ci gaba shafi tare da kasuwanci Ascentis Express RP-Amide shafi, an lura da cewa rabuwa yi na PMP shafi ya kasance mafi girma ga rarrabuwar kayyade shafi na kasuwanci effici.
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, don haka bukatar tsarkakewa na peptides kira na chromat. na sunadarai a cikin ruwa na jiki, 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'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 nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i nau'i). na masu nazarin da ke shiga cikin ma'auni mai yawa a lokacin da aka ba da lokaci4,5,6. Bugu da ƙari, a lokacin rabuwa na ruwa, ana iya mayar da hankali 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 proteomic17,8,9.
Reversed-lokaci ruwa chromatography (RP-LC) ne yadu amfani da tsarkakewa da kuma rabuwa da peptide gaurayawan ta yin amfani da octadecyl-gyara silica (ODS) a matsayin m phase11,12,13.Duk da haka, RP tsayayyun lokuta ba su samar da m rabuwa da peptides da sunadarai saboda su hadaddun tsarin,1 amphire tsari da kuma 1 amphire tsarin. Ana buƙatar matakan tsayuwar da aka tsara don nazarin peptides da sunadarai tare da nau'in polar da wadanda ba na iyakacin duniya ba 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.An yi amfani da ginshiƙai da yawa da aka yi amfani da su tare da lokaci-lokaci tare da ma'auni. Peptide da rarrabuwar fursunoni17,28,19,21.Mixed-Yanayin Kudi na (Wax) sun dace da gaban Peplide da RPLC), HPLC), HPLC Mai amfani da tsawan tsakaitattun abubuwa tare da Ƙungiyoyin polar da ke da alaƙa da haɗin gwiwa suna nuna kyakkyawan ikon rabuwa da zaɓi na musamman don ƙididdiga na iyakacin duniya 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 lokaci mai tsayayye na dodecyl-terminated polyamine kuma ya sami nasarar raba hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, da sauran masu nazari. ginshiƙai) ana samunsu ta kasuwanci a ƙarƙashin sunan kasuwanci Ascentis Express RP-Amide ginshiƙan, amma ana amfani da waɗannan ginshiƙan don nazarin amine 33 kawai.
A cikin binciken na yanzu, an shirya wani lokaci mai tsayi mai tsayi (N-phenylmaleimide-embedded polystyrene) kuma an kimanta shi don rabuwa da peptides da trypsin digests na HSA. An shirya lokaci na tsaye ta amfani da dabarun da ke gaba. An shirya ƙwayoyin silica na porous bisa ga tsarin da aka ba a cikin littafinmu na baya tare da wasu gyare-gyare na gyare-gyare na glycol. (PEG), TMOS, 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 su derivatize silica barbashi don shirya wani iyakacin duniya saka a tsaye phase.Sakamakon tsayawa idãnun lokaci da aka columnized a cikin wani bakin karfe 1.0. Shirye-shiryen shiryawa.An taimaka marufi na ginshiƙi tare da girgizar injiniya don tabbatar da an kafa gado mai kama da juna a cikin ginshiƙi.Yi la'akari da rabewar ginshiƙi mai cike da cakuda peptide wanda ya ƙunshi peptides biyar; (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) da Trypsin digest na ɗan adam serum albumin (HAS) .A peptide cakuda da trypsin digest na HSA aka lura raba tare da mai kyau ƙuduri da kuma yadda ya dace da PMP aiki idan aka kwatanta da cewa separation da aka kwatanta. Rukunin RP-Amide.Dukansu peptides da furotin an lura da su da kyau da kuma inganci a kan 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), Styroxy-H4, Benzol, Styroxy-H4. Peroxide (BPO), 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 ice-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 kashe a cikin wani tabo. an bushe shi a 70 ° C na tsawon sa'o'i 12. Busasshen 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 farfajiya.
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, kuma 0.1 ml na 2,2′-azoisobutyronitrile (AIBN) aka kara zuwa dauki flask don shirya phenylmaleimide-methyl vinyl isocyanate copolymer. tace kuma a bushe a cikin 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 faduwa mazurari.The cakuda aka refluxed a 180 hours tace da wash. 60 ° C na 3 hours. Sa'an nan kuma, PMCP-bonded silica barbashi (100 g) an narkar da a toluene (200 ml) da kuma 4-hydroxy-TEMPO (2 mL) da aka kara a gaban 100 µL na dibutyltin dilaurate a matsayin mai kara kuzari. An zuga cakuda a 80 ° C da aka bushe a 50 ° C.
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. (Bare silica and 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 suturar sputter Q150T, kuma an ajiye wani Layer na 5 nm Au a kan samfurori. Wannan yana inganta ingantaccen tsari ta amfani da ƙananan ƙarfin lantarki kuma yana samar da hatsi mai kyau, sanyi spultham, USA Electron. EA1112 elemental analyzer da aka yi amfani da elemental analysis.A Malvern (Worcestershire, UK) Mastersizer 2000 barbashi size analyzer da aka yi amfani da su don samun barbashi girman rabo. benci na Mastersizer.An gudanar da bincike na Thermogravimetric a cikin adadin 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-liyi, 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 wani tsayayye lokaci slurry ta dakatar da 150 MG na columnar mL da kuma aika shi zuwa 1 metan. An yi amfani da shi azaman slurry da sauran ƙarfi. Cika shafi a jere ta hanyar yin amfani da matsa lamba na 100 MP na minti 10, 80 MP na minti 15, da 60 MP na minti 30. A lokacin shiryawa, an yi amfani da vibration na inji tare da biyu GC shafi shakers (Alltech, Deerfield, tabbatar da sakin layi na IL, Amurka) matsa lamba sannu a hankali don hana duk wani lalacewa a cikin ginshiƙi.Cire haɗin ginshiƙi daga na'urar 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 musamman µLC na'urar ganowa (UV-2075) da gilashin-lined ƙuƙuntacciyar ƙuƙumman tubing da karin tasiri. shafi band broadening.Bayan marufi, capillaries (50 μm id 365 da rage union capillaries (50 μm) aka shigar a 1/16 ″ kanti na rage ƙungiyar. Ana yin tattara bayanai da sarrafa chromatographic ta amfani da Multichro 2000 software. Kulawa a 254 nm Analytes da aka gwada ta hanyar Chromat Analytes. (Northampton, MA).
Albumin daga jini na 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 na 37 ° C. 0.1% TFA.Tace maganin kuma adana ƙasa da 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 .SEM Hotuna na silica barbashi danda (A, B) sun nuna cewa, da bambanci da mu na baya karatu, wadannan barbashi ne mai siffar zobe a cikin abin da barbashi da aka elongated ko da rashin daidaituwa daidaita.The surface na ligand-bonded silica barbashi (C, D) shi ne santsi fiye da na danda silica zama mai santsi a kan rufi na polyethylene saboda da rufi na polyethylene. silica barbashi.
Ana duba hotunan siliki na siliki (A, B) da ligand-bonded silica (C, D).
The barbashi size rabawa na danda silica barbashi da ligand-bonded silica barbashi an nuna a cikin Figure 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 na baya binciken da aka kwatanta a cikin 5 The size of silica (5). na PMP ne 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 batch yana da narrower barbashi size rarraba idan aka kwatanta da mu baya binciken saboda sãɓãwar launukansa rabo na PEG, urea, TMOS, da kuma acetic acid part a cikin cakuda da ya fi girma girma a cikin lokaci na cakuda. polystyrene-daure silica barbashi lokaci da muka yi nazari a baya.Wannan yana nufin cewa surface functionalization na silica barbashi da styrene kawai ajiye polystyrene Layer (0.97 µm) a kan silica surface, alhãli kuwa a cikin PMP lokaci Layer kauri ya 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 kuma surface area na silica barbashi na 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 cikin Figure 3 (B) .Sakamako ne m ga mu baya binciken.The pore masu girma dabam na danda da ligand-1 sashi da 3 silica 3 particular, 1, 1, 1, 1 da silica. yana nuna cewa girman pore yana raguwa da 69 bayan gyare-gyaren sinadarai, kamar yadda aka nuna a cikin Table 1 (B), kuma an nuna canjin yanayin a cikin siffa 3 (B). binciken mu na baya (124 m2 / 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 kuma 10.21%, bi da bi) 42. The carbon loading na yanzu a cikin Bugu da kari ga SP lokaci a cikin lokaci mai tsawo, saboda SP. styrene, wasu polar 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. phenylmaleimide.Hakazalika, abubuwan da aka yi amfani da carbon na samfurori (4) da (5) sun kasance 2.7% da 2.9%, bi da bi, yayin da nauyin carbon na samfurin ƙarshe (6) ya kasance 6.35%, kamar yadda aka nuna a cikin Table 2. An duba hasara mai nauyi tare da PMP tsaye lokaci, kuma an nuna TGA mai lankwasa a cikin adadi mai kyau na 4. TGA mai lankwasa yana nuna alamar 8. da carbon loading (6.35%) saboda ligands ƙunshi ba kawai C amma kuma N, O, da H.
The phenylmaleimide-methylvinylisocyanate ligand da 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 kungiyar da ke da matsakaicin hulda tsakanin analy tashoshi da kuma babu wani matsakaicin hulda tsakanin analy tasha da analy. lokaci, tun da phenylmaleimide moiety ba shi da kama-da-wane cajin a al'ada pH.The polarity na a tsaye lokaci za a iya sarrafawa da mafi kyau duka adadin styrene da dauki lokaci na free radical polymerization.The karshe mataki na dauki (free-m polymerization) ne m da kuma iya canza polarity na a tsaye lokaci.Elemental bincike da aka lura da wadannan lokaci na loading lokaci da aka lura da lokacin da loading lokaci. kuma lokacin amsawa ya karu da ɗaukar nauyin carbon na lokaci mai tsayi kuma mataimakin versa.SPs da aka shirya tare da nau'i daban-daban na styrene suna da nauyin nauyin carbon daban-daban. Bugu da ƙari, ɗora waɗannan matakan tsaye a cikin ginshiƙan bakin karfe da kuma duba aikin su na chromatographic (zaɓi, ƙuduri, N darajar, da dai sauransu) .Bisa akan waɗannan gwaje-gwajen, an zaɓi tsarin da aka gyara don tabbatar da tsarin lokaci mai kyau na PMP don tabbatar da lokaci mai kyau na lokaci-lokaci mai sarrafawa na PMP.
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 P) yana ba da ƙimar 3 / m P. Ana nuna ginshiƙai da chromatograms 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 sun kasance masu kyau sosai, 13,500 ± 330 a kowace shafi (100 × 1.8 mm id), Daidai ne ga 135 / 00 Fig. ginshiƙai (100 × 1.8 mm id) an cika su da nau'ikan nau'ikan nau'ikan nau'ikan nau'ikan PMP guda uku don bincika haɓakawa.An yi rikodin ƙaddamarwar nazari ga kowane shafi ta amfani da yanayin haɓaka mafi kyau da adadin faranti na ka'idar N da lokacin riƙewa don raba cakuda gwajin guda ɗaya akan kowane ginshiƙi.An nuna bayanan sake fasalin ga ginshiƙan PMP a cikin ginshiƙan haɓakawa sosai a cikin Table 4. % Ƙimar RSD, kamar yadda aka nuna a Table 3.
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 chromatography na ruwa. chromatogram a cikin Hoto 6 ya nuna cewa samfurin ya rabu da kyau kuma ƙuduri yana da kyau sosai. An bincikar HSA diests ta amfani da ƙimar 100 µL / min, motsi na ruwa na 100 µL / min, lokaci na 3 na ruwa da 70 / 0ni. TFA.Kamar yadda aka nuna a cikin 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 layin layi na lokaci na wayar hannu. Ƙaƙwalwar ginshiƙan PMP shine 2.5 × 10-14 m2, yawan gudu ya kasance 25 μL / min, da 60/40 v / v ACN / ruwa da aka yi amfani da 1 mm permeability. id) yayi kama da na binciken mu na baya Ref.34.The permeability na ginshiƙi cushe da superficially 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 . barbashi Domin 5 μm barbashi 43. Saboda haka, da permeability na PMP lokaci ne kama da na 5 μm core-harsashi barbashi.
inda Wx shine nauyin ginshiƙan da aka cika da chloroform, Wy shine nauyin ginshiƙi wanda 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 (1) 0.8 mm ginshikan ginshikan 3 × 1 da 80 ginshikan. C18-Urea 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 tsayayyen lokaci. Silica na Silica saboda a cikin C18-Stenauki Files na C18 Ligands an haɗa su da silica mai tsayi, a cikin Polystyrene Plains.
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. An shirya cakuda peptide da aka zaɓa (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) a cikin 20 µL shafi da kuma Ascentis Express RP-Amide shafi sun kasance 2.6 µm da 3.9 µm, bi da bi. The HETP dabi'u nuna cewa rabuwa yadda ya dace na PMP shafi (100 × 1.8 mm id) ya fi kyau fiye da kasuwanci samuwa Ascentis Express RP-Amide shafi (100 × 1. Fig.) cewa raguwa a cikin darajar N tare da karuwa mai girma ba shi da mahimmanci idan aka kwatanta da bincikenmu na baya. Mafi girman haɓakar rabuwa na ginshiƙi na PMP (100 × 1.8 mm id) idan aka kwatanta da Ascentis Express RP-Amide shafi yana 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). ACN/H2O tare da 0.1% TFA.
A iyakacin duniya-ebedded polystyrene m lokaci da aka shirya da 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 ginshikan aiki da irin wannan size da kuma girman da PMP. Girman silica barbashi, sarrafawa kira na m 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 na daban-daban sunadaran, mun yi amfani da ginshikan na halitta fili daga cikin halitta fili amfani da ginshikan gina jiki. shuke-shuken magani da kayan aikin fungal a cikin chromatography na ruwa.A nan gaba, ginshiƙan PMP kuma za a kimanta don rabuwa da sunadarai da ƙwayoyin rigakafi na monoclonal.
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