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Qaybaha silica ee daloola ayaa lagu diyaariyey habka sol-gel iyadoo qaar ka mid ah wax laga beddelayo si loo helo qaybo makroporous ah. Qaybahaas waxaa laga soo saaray by wareejinta silsiladda jajabinta isku-darka dheeriga ah (RAFT) polymerization with N-phenylmaleimide-methylvinylisocyanate (PMI) iyo styrene si ay u diyaariyaan N-phenylmaleimiderid-polymstyrerrow stadium (PMI) 100 × 1.8 mm id) waxaa lagu rakibay baakad dhoobo ah.Qiimeynta PMP tiirka kala soocida isku dhafka peptide ka kooban shan peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin albamka) Xaaladaha aan fiicneyn, tirada saxanada aragtida ee isku dhafka peptide waa mid aad u sarreeya sida 280,000 taarikada / m². Marka la barbardhigo waxqabadka kala soocida tiirka la horumariyay ee ganacsiga Ascentis Express RP-Amide column, waxaa la arkay in waxqabadka kala soocida tiirka PMP uu ka sarreeyay tiirka ganacsiga marka loo eego hufnaanta iyo xallinta kala-soocidda.
Sanadihii la soo dhaafay, warshadaha biopharmaceutical waxay noqdeen suuq caalami ah oo sii fidaya iyadoo koror la taaban karo uu ku yimid saamiga suuqa dareeraha, unugyada iyo unugyada waa hawl aad u adag sababtoo ah tirada badan ee noocyada suurtagalka ah ee la ogaan karo ee hal muunad.In kasta oo spectrometry mass ay tahay qalab wax ku ool ah oo loogu talagalay peptide iyo isku xigxiga borotiinka, haddii shaybaarada noocan oo kale ah lagu duro spectrometer mass in hal pass, kala soociddu ma noqon doonto mid ku habboon. Dhibaatadani LC waa la yarayn karaa iyada oo la fulinayo nambarada chromatography ee ka hor inta aan la soo saarin falanqaynta MS. spectrometer waqti la siiyay4,5,6.Intaa waxaa dheer, inta lagu jiro kala-soocidda wajiga dareeraha, falanqeeyayaashu waxay diiradda saari karaan gobollada cidhiidhiga ah, taas oo xoogga saaraysa falanqaynta iyo hagaajinta dareenka MS. Dareeraha chromatography (LC) ayaa si weyn u horumaray tobankii sano ee la soo dhaafay waxayna noqotay farsamo caan ah oo ku saabsan falanqaynta borotiinka7,8,9,10.
Dib-u-celinta dareeraha chromatography (RP-LC) ayaa si weyn loogu isticmaalaa nadiifinta iyo kala-soocidda isku-dhafka peptide-ka iyadoo la adeegsanayo octadecyl-modified silica (ODS) sida marxaladda taagan waxaa looga baahan yahay in lagu falanqeeyo peptides iyo borotiinada leh qaybaha polar iyo kuwa aan cirridka ahayn si ay ula falgalaan oo ay u sii hayaan analytes16. Isku-dhafka isku-dhafka ah ee chromatography, kaas oo bixiya isdhexgalka multimodal, wuxuu noqon karaa beddelka RP-LC ee kala-soocidda peptides, borotiinnada, iyo isku-dhafka kale ee isku dhafan. 8,19,20,21.Mixed-mode taagan wejiyada (WAX / RPLC, HILIC / RPLC, polar intercalation / RPLC) waxay ku haboon yihiin peptide iyo kala-soocidda borotiinka sababtoo ah joogitaanka kooxaha polar iyo non-polar labadaba22,23,24,25,26,27,28 falanqeeyayaasha polar iyo kuwa aan dabaylaha ahayn, maadaama kala-saarku ku xidhan yahay isdhexgalka u dhexeeya falanqaynta iyo marxaladda taagan.Isdhexgalka Multimodal 29, 30, 31, 32. Dhawaan, Zhang et al.30 waxay diyaarisay dodecyl-la joojiyay marxaladda taagan ee polyamine waxayna si guul leh u kala soocday hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, iyo dhowr analytes kale. Waxay ku jirtaa magaca ganacsiga Ascentis Express RP-Amide columns, laakiin tiirarkan waxaa loo isticmaalaa falanqaynta amin 33 oo keliya.
Daraasadda hadda jirta, marxaladda taagan ee cirifka ah (N-phenylmaleimide-embedded polystyrene) ayaa la diyaariyey oo lagu qiimeeyay kala-soocidda peptides iyo trypsin dheefshiidka ee HSA. Wajiga taagan ayaa la diyaariyey iyadoo la adeegsanayo istaraatiijiyada soo socota. Qaybaha silica ee porous ayaa loo diyaariyey iyadoo la raacayo habraaca lagu bixiyay daabacaaddeenii hore iyada oo la beddelayo qaar ka mid ah diyaarinta glycol ee glycol. OS, acetic acid biyaha ayaa la hagaajiyay si loo diyaariyo qaybo silica leh size pore weyn.Labaad, ligand cusub, phenylmaleimide-methyl vinyl isocyanate, ayaa la farsameeyey oo loo isticmaalo in derivatize qaybaha silica si ay u diyaariyaan a polar ah gundhig taagan stage.The natiijada taagan ee marxaladda taagan waxaa lagu soo buuxiyey galay a ahama columned line 1.0.8. oo leh gariir farsamo si loo hubiyo in sariir isku mid ah laga sameeyay tiirka dhexdiisa(Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) iyo Trypsin dheefshiidka ee serum albumin aadanaha (HAS) . Isku darka peptide iyo dheefshiidka trypsin ee HSA ayaa lagu arkay si ay u kala soocaan xallinta wanaagsan iyo hufnaan ee PRP. Labada peptides iyo borotiinada labadaba waxaa loo arkay inay si fiican u xalliyeen oo ay waxtar u leeyihiin tiirka PMP, kaas oo ka waxtar badnaa tiirka Ascentis Express RP-Amide.
PEG (Polyethylene Glycol), Urea, Acetic Acid, Trimethoxy Orthosilicate (TMOS), Trimethoxy Chlorosilane (TMCS), Trypsin, Serum Albumin (HSA), Ammonium Chloride, Urea, Hexane Methyldisilazane (HMDS), Methacryloyl Chloride (Mc), Peroxide, Benzoroxy-PerydOxide (MC), HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, iyo Acetone Laga soo iibsaday Sigma-Aldrich (St. Louis, MO, USA).
Isku dar ah HOL (8 g), polyetylen glycol (8 g), iyo 8 mL oo ah 0.01 N acetic acid ayaa la walaaqay 10 daqiiqo, ka dibna 24 mL oo TMOS ah ayaa lagu daray xaaladaha baraf-qabow. Isku-darka falcelinta waxaa lagu kululeeyay 40 ° C 6 saacadood ka dibna 120 ° C biyo yar ayaa lagu shubay 8 saacadood oo biyo ah. 0 ° C saacadaha 12. Cufka jilicsan ee la qalajiyey ayaa ahaa dhul siman oo foorno ah oo lagu dhajiyay 550 ° C saacadaha 12. Saddex qaybood ayaa la diyaariyey oo lagu gartaa in lagu baaro dib-u-soo-nooleynta cabbirka qaybaha, xajmiga daloolka iyo aagga dusha sare.
Beddelka dusha sare ee qaybaha silica leh ligand phenylmaleimide-methylvinylisocyanate (PCMP) oo ay ku xigto radial polymerization oo leh styrene, koox koox-kooxeed ah ayaa la diyaariyey.Marxaladda taagan ee isku-darka iyo silsiladaha polystyrene. Habka diyaarinta ayaa lagu qeexay hoos.
N-phenylmaleimide (200 mg) iyo methyl vinyl isocyanate (100 mg) ayaa lagu milmay toluene qalalan, iyo 0.1 mL oo ah 2,2′-azoisobutyronitrile (AIBN) ayaa lagu daray weelka falcelinta si loogu diyaariyo phenylmaleimide-methyl vinyl isocyanate copolymer foornada 40 ° C ilaa 3 saacadood.
Qaybaha silica ee la qalajiyey (2 g) ayaa lagu kala firdhiyey toluene qalalan (100 mL), walaaqay oo lagu sonicated 500 mL wareega hoose ee wareega 10 min.PMCP (10 mg) ayaa lagu milmay toluene waxaana lagu daray dropwise in flask falcelinta iyada oo loo marayo funnel hoos u dhac. Saacadaha 3. Kadibna, qaybaha silica ee PMCP-ku-xidhan (100 g) ayaa lagu milmay toluene (200 ml) iyo 4-hydroxy-TEMPO (2 mL) ayaa lagu daray iyadoo ay joogaan 100 µL of dibutyltin dilaure sida kicinta. Isku-darka waxaa lagu walaaqay 50 ° C oo la qalajiyey saacadaha 8, 5 ° C.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), iyo TEMPO-PMCP-ku-xidhan qaybaha silica (1.5 g) ayaa lagu kala firdhiyey toluene waxaana lagu nadiifiyaa nitrogen.
Shaybaarada ayaa lagu dhajiyay 393 K saacada 1 si loo helo cadaadiska hadhaaga ah ee ka yar 10-3 Torr. Qadarka N2 ee lagu dhajiyay cadaadiska qaraabada ah ee P / P0 = 0.99 ayaa loo isticmaalay si loo go'aamiyo wadarta guud ee mugga daloolka. The morphology of bare and ligand-bonded silica particles ayaa lagu baaray shaybaarka elektaroonigga ah ee Japan (High microscori Japan). Qaybaha silica ligand-bonded) ayaa lagu dhejiyay tiir aluminium ah iyadoo la adeegsanayo cajalad kaarboon koolo ah.Gold ayaa lagu dhejiyay shaybaarrada iyadoo la adeegsanayo koodhka Q150T, iyo lakabka 5 nm Au ayaa lagu shubay shaybaarada .A Malvern (Worcestershire, UK) Mastersizer 2000 falanqeeye cabbirka qayb ka mid ah ayaa loo isticmaalay si loo helo qaybinta qaybinta qaybinta. 5 °C daqiiqadii in ka badan heerkul u dhexeeya 30 ilaa 800 °C.
Dhalada ku dahaaran tiirarka cidhiidhiga ah ee birta dhuuban ee cabbirka (100 × 1.8 mm id) ayaa la soo buuxiyay iyadoo la adeegsanayo habka xirxirida qulqulka, iyadoo la adeegsanayo isla nidaamka loo adeegsaday Ref.31.A tiir bir ah oo bir ah (glass-lined, 100 × 1.8 mm id) oo leh meel bannaan oo ka kooban 1 µm frit ayaa lagu xiray baakadaha slurry (Alltech Deerfield, IL, USA) . Diyaari slurry wejiga taagan adigoo joojinaya 150 mg ee column column ee loo isticmaalo methan 2. dareere slurry iyo sidoo kale dareeraha kicinta. Buuxi tiirka si isdaba joog ah adoo isticmaalaya cadaadiska 100 MP ee 10 daqiiqo, 80 MP ee 15 daqiiqo, iyo 60 MP ee 30 minutes. Inta lagu jiro xiritaanka, gariir farsamo ayaa lagu dabaqay laba ruxood oo GC ah (Alltech, Deerfield, IL, USA) si loo hubiyo in si tartiib tartiib ah loo sii daayo tiirarka khariidadaha. Xidhiidhka ka saar tiirka cutubka baakadaha dareeraha ah oo ku xidh mid kale oo ku habboon meesha laga soo galo iyo nidaamka LC si aad u hubiso waxqabadkiisa.
Bamka LC (10AD Shimadzu, Japan), injector (Valco (USA) C14 W.05) oo leh 50nL duritaanka loop, xuubka degasser (Shimadzu DGU-14A), daaqada capillary UV-VIS waxaa la dhisay Gaarka ah µLC qalab detector (UV-2075) iyo xidhidha dhalooyinka-lidan tubbada xidhmooyin aad u gaaban. Baakadaha ka dib, capillaries (50 μm id 365 iyo yaraynta capillaries midowga (50 μm) ayaa lagu rakibay 1/16 "outlet ee ururka dhimista. Xog ururinta iyo habaynta chromatographic waxaa la sameeyey iyada oo la isticmaalayo software Multichro 2000. Kormeerka at 254 nm Oriented Analytes ayaa lagu tijaabiyey UVginography, MAChromat data.
Albumin laga bilaabo serum bini'aadamka, budada lyophilized, ≥ 96% (agarose gel electrophoresis) 3 mg oo lagu qaso trypsin (1.5 mg), 4.0 M urea (1 ml), iyo 0.2 M ammonium bicarbonate (1 mL) xalinta beddel oo ku kaydi wax ka hooseeya 4 °C.
Kala soocida isku dhafka peptide iyo dheefshiidka HSA trypsin ayaa si gaar ah loogu qiimeeyay tiirarka PMP. Hubi kala soocida isku dhafka peptide iyo dheefshiidka trypsin ee HSA ee tiirka PMP oo barbar dhig natiijada safka Ascentis Express RP-Amide. Tirada saxanka aragtida waxaa loo xisaabiyaa sida soo socota:
Sawirada SEM ee qaybaha silica qaawan iyo qaybaha silica ee isku xidhan ayaa lagu muujiyay FIG.2 .Saam Sawirada ee loo yaqaan 'Capicals'
Sawirada mikroskoob ee elektarooniga ah ee walxaha silica qaawan (A, B) iyo qaybaha silica-ku-xidhan ee isku xidhan (C, D).
Qaybinta cabbirka qaybinta qaybaha silica qaawan iyo qaybaha silica ee ligand-ku-xidhan ayaa lagu muujiyay Jaantuska 3 (A) . Qaybinta qaybinta qaybinta qaybinta mugga ku salaysan waxay muujisay in cabbirka qaybaha silica uu kordhay ka dib markii wax laga beddelo kiimikaad (Sawir 3A) . 3.36 μm, marka la barbar dhigo daraasaddeena hore ee ad (0.5) qiimaha 3.05 μm (polystyrene-bound silica particles) 34. Dufcaddani waxay lahayd qaybinta qaybinta qayb yar oo cidhiidhi ah marka loo eego daraasaddeena hore sababtoo ah saamiga kala duwan ee PEG, HOL, TMOS, iyo acetic acid ee qaybta falcelinta ee qayb ka mid ah qayb ka mid ah qaybta MP si ka weyn qaybta PEG. Horey loo bartay
Qaybinta cabbirka walxaha (A) iyo qaybinta cabbirka daloolka (B) ee qaybinta silica qaawan iyo qaybaha silica-ku-xidhan ee ligand.
Cabbirka daloolada, mugga daloolka iyo dusha sare ee qaybaha silica ee daraasadda hadda jirta ayaa lagu bixiyaa Shaxda 1 (B) . Noocyada PSD ee qaybaha silica ee qaawan iyo ligand-bonded silica particles ayaa lagu muujiyay sawirka 3 (B) Natiijadu waxay u dhigantaa daraasaddeenii hore cabbirka re wuxuu hoos u dhacayaa 69 ka dib markii wax laga beddelay kiimikaad, sida ku cad Shaxda 1 (B), iyo isbeddelka qalooca ayaa lagu muujiyay sawirka 3 (B) .Si la mid ah, mugga daloolka ee qaybaha silica ayaa hoos u dhacay 0.67 ilaa 0.58 cm3 / g ka dib markii wax laga beddelo kiimikada g) .Sida ku cad Jadwalka 1 (B), aagga dusha sare (m2 / g) ee qaybaha silica ayaa sidoo kale hoos u dhacay 116 m2 / g ilaa 105 m2 / g ka dib isbeddelka kiimikada.
Natiijooyinka falanqaynta elemental ee marxaladda taagan ayaa lagu muujiyay shaxda 2. Xamuulka kaarboon ee marxaladda taagan ee hadda taagan waa 6.35%, taas oo ka hooseysa culeyska kaarboon ee daraasaddeena hore ( polystyrene bonded silica particles, 7.93% 35 iyo 10.21%, siday u kala horreeyaan) ligands sida phenylmaleimide-methylvinylisocyanate (PCMP) iyo 4-hydroxy-TEMPO ayaa loo isticmaalay.Miisaanka nitrogen ee boqolkiiba marxaladda taagan hadda waa 2.21%, marka la barbar dhigo 0.1735 iyo 0.85% by miisaanka nitrogen ee daraasadihii hore, siday u kala horreeyaan.Tani waxay ka dhigan tahay in wt % of carbon load marxaladda hadda jira ay sabab u tahay. 4) iyo (5) waxay ahaayeen 2.7% iyo 2.9%, siday u kala horreeyaan, halka culeyska kaarboonka ee alaabta kama dambaysta ah (6) uu ahaa 6.35%, sida ku cad Jadwalka 2. Miisaanka dhimista waxaa lagu hubiyay marxaladda taagan ee PMP, iyo TGA-ga ayaa lagu muujiyay Jaantus 4. TGA TGA waxay muujinaysaa miisaan lumis ah 8.6% , laakiin ligands ayaa sidoo kale ku jira boqolkiiba C. iyo H.
The phenylmaleimide-methylvinylisocyanate ligand ayaa loo doortay wax ka beddelka dusha sare ee qaybaha silica sababtoo ah waxay leedahay kooxaha phenylmaleimide polar iyo kooxaha vinylosyanate.Vinyl isocyanate group.Vinyl isocyanate group.Vinyl isocyanate groups more react with styrene by live radical polymerization.Sababta labaad waa in la geliyo koox leh isdhexgalka dhexdhexaadka ah iyo analy ee saldhigga analy ah. Nylmaleimide moiety ma laha lacag dalwaddii pH caadiga ah. The polarity ee marxaladda taagan waxaa lagu xakameyn karaa qadarka ugu fiican ee styrene iyo waqtiga falcelinta ee polymerization xagjirka lacag la'aan ah.The ugu dambeeya ee falcelinta (polymerization-free xagjirka ah) waa mid muhiim ah oo bedeli kara polarity ee wejiga taagan. Falanqaynta Elemental ayaa la sameeyay si loo hubiyo in kaarboon loading kaarboonka ee wakhtiga kor u kacay ee loading kuwaas oo la arkay. Mar labaad, ku shub wejiyadan taagan tiirarka birta ah ee aan lahayn iyo hubi waxqabadkooda chromatographic (doorashada, xallinta, qiimaha N, iwm.) Iyadoo lagu saleynayo tijaabooyinkan, qaabaynta la hagaajiyay ayaa loo doortay si loo diyaariyo marxaladda taagan ee PMP si loo hubiyo ilaalinta polarity iyo falanqaynta wanaagsan.
Shan isku dhaf ah oo peptide ah (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) ayaa sidoo kale lagu qiimeeyay iyadoo la adeegsanayo tiirka PMP iyadoo la adeegsanayo marxalad wareeg ah;60/40 (v/v) acetonitrile/biyaha (0.1% TFA) oo ah heerka socodka 80 μL/min. Marka la eego xaaladaha ugu fiican, lambarka saxanka aragtida (N) tiirkiiba (100 × 1.8 mm id) waa 20,000 ± 100 (200,000) ee chromium iyo 3 N. atograms ayaa lagu muujiyay Jaantuska 5A. Falanqaynta degdega ah ee tiirka PMP ee heerka socodka sare (700 μL / min), shan peptides ayaa lagu dhajiyay hal daqiiqo gudaheed, N qiyamka ayaa aad u wanaagsan, 13,500 ± 330 halkii tiir (100 × 1.8 mm id), Waxay u dhigantaa 135,000 saxan / m.8 mm id) waxaa lagu soo buuxiyey saddex qaybood oo kala duwan oo PMP ah oo taagan si loo hubiyo dib-u-soo-nooleynta. Falanqaynta falanqaynta ee tiir kasta ayaa la duubay iyadoo la adeegsanayo xaaladaha ugu fiicnaanta iyo tirada taarikada aragtida N iyo wakhtiga haynta si loo kala saaro isku dhafka tijaabada ah ee tiir kasta.
Kala soocida isku dhafka peptide ee tiirka PMP (B) iyo Ascentis Express RP-Amide column (A);wajiga mobaylka 60/40 ACN/H2O (TFA 0.1%), cabbirada tiirka PMP (100 × 1.8 mm id);Analytical Nidaamka elution ee xeryahooda: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) iyo 5 (leucine) acid enkephalin)).
Tiirka PMP (100 × 1.8 mm id) ayaa lagu qiimeeyay kala soocida dheefshiidka tryptic ee serum albumin ee waxqabadka sare ee chromatography dareeraha. chromatogram (Jaantus 6), dheef-shiid kiimikaadka HSA ayaa loo kala qaybiyay 17 ugu sarreeya oo u dhiganta 17 peptides. Waxtarka kala-soocidda ee mid kasta oo ugu sarreeya ee dheef-shiid kiimikaadka HSA ayaa la xisaabiyay waxaana qiimaha lagu bixiyaa shaxda 5.
Dheefshiidka tijaabinta ee HSA (100 × 1.8 mm id) ayaa lagu kala soocay tiirka PMP;heerka socodka (100 µL/min), wareega wareega 60/40 acetonitrile/biyo leh 0.1% TFA.
halka L waa dhererka tiirarka, η waa viscosity ee wajiga wareega, ΔP waa cadaadiska dhabarka ee tiirka, iyo u waa xawaaraha toosan ee wajiga moobiilka. Dareemida tiirka PMP wuxuu ahaa 2.5 × 10-14 m2, heerka qulqulka wuxuu ahaa 25 μL / min, iyo 60/40 v / v ACN / biyaha ayaa la mid ah 1 × 0. Midda daraasaddeennii hore ee Ref.34. Dareemida tiirka ay ka buuxaan qaybo dusha sare ka daloola waa: 1.7 × 10-15 ee 1.3 μm qaybo, 3.1 × 10-15 oo ah 1.7 μm qaybo, 5.2 × 10-15 iyo 2.5 μm 2.5 m2. 43.Sidaas darteed, daah-furnaanta wajiga PMP waxay la mid tahay 5 μm qaybaha qolofka xudunta u ah.
halka Wx uu yahay culeyska tiirka lagu kariyey chloroform, Wy waa culeyska tiirka ka buuxa methanol, iyo ρ waa cufnaanta dareeraha. Cufnaanta methanol (ρ = 0.7866) iyo chloroform (ρ = 1.484) .Guud ahaan porosity ee SILICA PARTICLES1 × 8 × 1.0.8 Tiirarka rea ​​31 ee aan hore u barannay waxay ahaayeen 0.63 iyo 0.55, siday u kala horreeyaan Nooca joogtada ah ee C18 ligands waxay ku xiran yihiin qaybaha silica sida silsilado toosan, halka marxaladaha taagan ee nooca polystyrene, lakabka polymer qaro weyn ayaa laga sameeyay hareeraha.
Jaantuska 7A,B wuxuu muujinayaa tiirka PMP (100 × 1.8 mm id) iyo Ascentis Express RP-Amide tiirka (100 × 1.8 mm id) iyadoo la adeegsanayo xaalado isku mid ah (ie, 60/40 ACN/H2O iyo 0.1% TFA).) ee shirqoolkii van Deemter.Isku darka peptide ee la xushay (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) ayaa lagu diyaariyay 20 µL/ Heerka socodka ugu yar ee labada tiir waa 800 µL/min. Heerka ugu hooseeya ee HETP waa Express RP-Amide column waxay ahaayeen 2.6 µm iyo 3.9 µm, siday u kala horreeyaan.Qiimaha HETP waxay muujinayaan in wax ku oolnimada kala-soocidda tiirka PMP (100 × 1.8 mm id) uu aad uga fiican yahay ganacsiga la heli karo Ascentis Express RP-Amide (100 × 1.8 mm) oo muujinaya hoos u dhac ku yimaadda shaxanka. Ma aha mid muhiim ah marka loo eego daraasaddeennii hore. Waxtarka sare ee kala-soocidda ee tiirka PMP (100 × 1.8 mm id) marka la barbar dhigo tiirka Ascentis Express RP-Amide wuxuu ku salaysan yahay hagaajinta qaabka qaybta, cabbirka, iyo hababka xirmooyinka adag ee tiirarka loo isticmaalo shaqada hadda34.
(A). 0.1% TFA.
A polar-embedded polystyrene stage stationary ayaa la diyaariyey oo lagu qiimeeyay kala soocida isku dhafka peptide synthetic iyo trypsin dheefshiidka ee serum albumin aadanaha (HAS) ee waxqabadka sare ee chromatography dareere ah. Qaybaha a, kontaroolo kontoroolka marxaladda taagan, iyo packing.In dheer in waxtarka sare ee kala soocidda column, hooseeyo cadaadiska dhabarka ee heerka socodka sare waa faa'iido kale ee this phase.PMP tiirarka taagan soo bandhigaan reproducibility wanaagsan oo loo isticmaali karaa falanqaynta isku dhafka peptide iyo dheefshiidka trypsin ee borotiinada kala duwan. Waxaan ku talo jirnaa in la isticmaalo tiirarka fungal ee dhirta dabiiciga ah ee ka soo xeryahooda dareere ah ee dhirta dabiiciga ah. chromatography. Mustaqbalka, tiirarka PMP ayaa sidoo kale lagu qiimeyn doonaa kala-soocidda borotiinka iyo unugyada unugyada monoclonal.
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