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鍩哄洜娌荤枟鏄敓鐗╁埗鑽涓氫腑涓€涓揩閫熷闀跨殑棰嗗煙锛岄€氳繃鍩哄洜娌荤枟鍙疄鐜扮柧鐥呯殑娌荤枟鎴栭闃层€傚叾涓紝閲嶇粍鑵虹浉鍏崇梾姣?rAAV)鏄洰鍓嶅熀鍥犳不鐤楅鍩熺爺绌惰緝澶氱殑涓€绫荤梾姣掕浇浣撱€侟/span>
鑵虹浉鍏崇梾姣掞紙adeno-associated virus, AAV锛夋槸寰皬鐥呮瘨绉戯紙Parvoviridae锛夊鏃忕殑鎴愬憳涔嬩竴锛屼竴鑸紝鐮旂┒涓噰鐢ㄧ殑閲嶇粍鑵虹浉鍏崇梾姣掕浇浣擄紙Recombination adeno-associated virus, rAAV锛夋槸鍦ㄩ潪鑷寸梾鐨勯噹鐢熷瀷AAV鍩虹涓婃敼閫犺€屾垚鐨勫熀鍥犺浇浣擄紝鐢变簬鍏剁绫诲鏍枫€佸厤鐤師鎬ф瀬浣庛€佸畨鍏ㄦ€ч珮銆佸涓荤粏鑳炶寖鍥村箍銆佹墿鏁h兘鍔涘己銆佷綋鍐呰〃杈惧熀鍥犳椂闂撮暱绛夛紝rAAV琚涓烘渶鏈夊墠閫旂殑鍩哄洜鐮旂┒鍜屽熀鍥犳不鐤楄浇浣撲箣涓€銆侟/span>
鐩墠锛宺AAV鐨勫噯纭畾閲忓垎鏋愬拰琛ㄥ緛鐨勯毦搴︽槸闃荤鍩哄洜娌荤枟蹇€熷彂灞曠殑鍏抽敭鍥犵礌銆傛垜浠父甯搁渶瑕佸rAAV杩涜缁煎悎鍏ㄩ潰琛ㄥ緛锛屾瘮濡傝。澹虫暟閲忋€佸疄蹇冪巼銆侀绮掑昂瀵搞€佽仛闆嗕綋姣斾緥绛夈€備紶缁熸儏鍐碉紝rAAV婊村害鍜岀梾姣掕浇閲忛噰鐢‥LISA銆乨dPCR銆丄UC鍜孍M绛夋妧鏈繘琛屾祴閲忋€備絾杩欎簺鏂规硶閫氬父璐规椂璐瑰姏锛岃€屼笖绮剧‘搴︿笉楂樸€侟/span>
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01浠櫒鍙傛暟
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涓庝紶缁烪PLC涓嶅悓锛屾祴閲忚繃绋嬩笉渚濊禆鏌变繚鐣欎綋绉紝涔熶笉闇€瑕佷竴绯诲垪鏍囨牱杩涜鑹茶氨鏌辨牎姝c€傚浘1鏄剧ず浜嗕娇鐢∣MNISEC娴嬮噺鐨凜QA鍏抽敭璐ㄩ噺鍙傛暟銆侟/span>


02 妫€娴嬫柟娉旤/strong>
鎴戜滑閲囩敤Empty鍜孎ull rAAV5涓や釜鏍峰搧浣滀负鍒嗘瀽妗堜緥銆侳ull rAAV5 杞芥湁宸茬煡鍒嗗瓙閲忎负785 kg/mol鐨凱FB-GFP ssDNA銆傜粡qPCR鍜孍LISA娴嬮噺鏂瑰紡鍙煡锛岃鏍锋湰鐨勭梾姣掓淮搴︿负2.5x1013銆侟/span>
閲囩敤鑹茶氨鏌盤4000鍜孭3000涓茶仈锛屽rAAV鏍峰搧鐨勮繘琛岃壊璋卞垎绂汇€傜敱OMNISEC杞欢閲囬泦鍒嗘瀽娴嬭瘯缁撴灉锛屽叾涓‖浠剁郴缁熷寘鍚玂MNISEC RESOLVE(鍖呭惈娉点€佽嚜鍔ㄨ繘鏍峰櫒鍜屾煴娓╃)鍜孫MNISEC REVEAL锛堝寘鍚ず宸€乁V/PDA鍜岀洿瑙?0掳/灏忚7掳鍏夋暎灏勬娴嬪櫒锛夈€侟/span>

鏍峰搧缁忚繃鍒嗙娲楄劚鍚庯紝浣跨敤鍏辫仛鐗╁垎鏋愭柟娉曠‘瀹氭牱鍝佷袱绉嶄笉鍚岀粍鍒嗙殑娴撳害鍜屽垎瀛愰噺銆傝绠楁柟娉曞涓嬶細

鍏朵腑锛孋oncCapsid鏄。澹虫祿搴?mg/mL)锛孨A鏄樋浼忎冀寰风綏鏁帮紝Mwcapsid鏄。澹崇殑鍒嗗瓙閲?g/mol)锛孋oncDNA鏄疍NA娴撳害(mg/mL)锛孧wSeqDNA鏄潵鑷簭鍒楃殑ssDNA鐨勫垎瀛愰噺銆傚洜姝わ紝閫氳繃璁$畻鍑虹殑棰楃矑娴撳害锛屽彲浠ュ緢瀹规槗鍦板緱鍑烘牱鍝佸疄蹇冪巼鐨勭櫨鍒嗘瘮銆侟/span>

03妫€娴嬬粨鏋淈/strong>
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鍗曚綋鐨勫垎瀛愰噺涓?.84脳106g/mol銆傝。澹崇殑鐞嗚鍒嗗瓙閲忎负3.8脳106g/mol锛岃瘉瀹炲垎鏋愮粨鏋滀笌棰勬湡鐩哥銆侟/span>
MW/Mn涓哄垎瀛愰噺鍒嗗竷锛屾弿杩颁簡鏍峰搧鐨勫垎鏁f€э紝鍗曚綋鍜屼簩鑱氫綋鐨勫€兼帴杩?锛岃€岃仛闆嗕綋鍜岀墖娈靛潎鏄剧潃楂樹簬1锛岃〃鏄庡湪鍚屼竴宄板唴鏈夊涓笉鍚屽垎瀛愰噺鐨勭粍鍒嗐€侟/span>
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Fraction of Protein鏄剧ず浜嗘牱鍝佷腑琛e3鐨勭櫨鍒嗘瘮锛屽崟浣撳寘鍚?9.8%鐨勮。澹炽€傝繖璇佸疄浜嗘牱鏈‘瀹炴槸Empty rAAV5銆侟/span>
鏈€鍚嶦mpty rAAV5鏍峰搧鎬绘淮搴︿负5.91x1013Vp/ml銆侟/span>

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瀵逛簬涓讳綋鐨勫崟浣撳嘲锛岃绠楀嚭鍏舵贩鍚堢墿鍒嗗瓙閲忎负4.49脳106g/mol锛屽叾涓?6%涓鸿。澹炽€俽AAV5鐨勮泲鐧借川缁勫垎鐨勫垎瀛愰噺涓?.89脳106g/mol锛岃繖涓庤〃1涓璄mpty rAAV5 鐨勬暟鎹竴鑷淬€傚崟浣撴槸鎬讳綋鐨?3.2%锛屾牱鏈殑鎬绘淮搴︿负7.48x1013VP/ml銆傚叾涓崟浣撳寘鍚?8% Full rAAV5锛?2% Empty rAAV5銆傞渶瑕佹敞鎰忕殑鏄紝杩欑鍒嗘瀽鏂规硶鍋囪鏍峰搧瑕佷箞鏄疐ull 锛岃涔堟槸Empty 锛屽拷鐣ラ儴鍒嗚杞芥垨杩囧害瑁呰浇鎯呭喌銆侟/span>
Zetasizer Ultra绾崇背绮掑害鍙婄數浣嶄华鍙互浣跨敤MADLS鏂瑰紡蹇€熺‘瀹氱梾姣掓淮搴︺€備粠OMNISEC鑾峰緱鐨勬暟鎹笌Zetasizer Ultra鐨勭矑瀛愭淮搴﹁繘琛屼簡姣旇緝锛屼袱绉嶆妧鏈箣闂存湁寰堝ソ鐨勭浉鍏虫€э紝瑙佸浘4銆侟/span>

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涓轰簡杩涗竴姝ヨ瘎浼癘MNISEC瀵箁AAV鏍峰搧鍑嗙‘琛ㄥ緛鑳藉姏锛屾垜浠繘琛屼簡rAAV5鏍峰搧鐨勭儹搴斿姏绋冲畾鎬х爺绌讹紝鍚屾椂锛屽熀浜嶼S Ultra瀵硅仛闆嗕綋鐨勬瀬楂樼伒鏁忓害锛屾垜浠埄鐢ㄤ簡ZS Ultra琛ㄥ緛rAAV5鑱氶泦浣撶殑寰皬鍙樺寲銆傛祴璇曟潯浠舵槸灏唕AAV5鏍峰搧缃簬25oC鍒?0oC涔嬮棿杩涜娴嬭瘯銆侟/span>

鍦ㄤ笉鏂姞鐑繃绋嬩腑锛屽湪姣忎釜娓╁害涓嬫祴閲弐AAV5鏍峰搧鐨勭矑寰勩€傚湪25oC鍜?5oC涔嬮棿锛屾病鏈夎瀵熷埌绮掑緞鐨勫彉鍖栥€備粠35oC寮€濮嬶紝鍙互瑙傚療鍒扮矑寰勫紑濮嬪澶э紝杩欒〃鏄庢牱鍝佸紑濮嬪彂鐢熷彉鍖?鍥?A)銆?0oC鍜?5oC涓嬬殑鏁版嵁姣旇緝娓呮鍦版樉绀轰簡杩欎簺鏍峰搧涔嬮棿鐨勫ぇ灏忓樊寮?鍥?B)銆傛垜浠€夋嫨45oC鏉′欢锛屽OMNISEC杩涜杩涗竴姝ョǔ瀹氭€х爺绌躲€侟/span>
灏唕AAV5鏍峰搧鍦ㄧǔ瀹氬湪45oC锛屽垎鍒湪2min 銆?min銆?0min鍜?5min鍚庯紝鍙栨牱鍝佸埌OMNISEC涓婃祴璇曘€傚浘7鑹茶氨鍙犲姞鍥炬樉绀烘牱鍝佸彂鐢熶簡鏄庢樉鐨勫彉鍖栵紝鑱氶泦浣撶櫨鍒嗗惈閲忓鍔狅紝鍗曚綋娴撳害鍚噺闄嶄綆銆傝〃4鏄剧ずMW鍦ㄦ娼滀紡鏈熷唴淇濇寔绋冲畾锛屽崟浣撳嘲涓殑AAV鐧惧垎姣斾篃淇濇寔绋冲畾銆侟/span>

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椹皵鏂囧笗绾崇 2021-12-27 | 闃呰锛?726

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