動物細胞大規模培養的生物反應器操作模式,一般分為分批式操作(batch)、流加式操作(Fed-batch)、半連續式操作(semi-continuous)、連續式操作(continuous)和灌流式操作(perfusion)五種操作模式。
1. 批式操作(batch culture)
批pi式shi操cao作zuo是shi動dong物wu細xi胞bao規gui模mo培pei養yang進jin程cheng中zhong較jiao早zao期qi采cai用yong的de方fang式shi,也ye是shi其qi它ta操cao作zuo方fang式shi的de基ji礎chu。該gai方fang式shi采cai用yong機ji械xie攪jiao拌ban式shi生sheng物wu反fan應ying器qi,將jiang細xi胞bao擴kuo大da培pei養yang後hou,一yi次ci性xing轉zhuan入ru生sheng物wu反fan應ying器qi內nei進jin行xing培pei養yang,在zai培pei養yang過guo程cheng中zhong其qi體ti積ji不bu變bian,不bu添tian加jia其qi它ta成cheng分fen,待dai細xi胞bao增zeng長chang和he產chan物wu形xing成cheng積ji累lei到dao適shi當dang的de時shi間jian,一yi次ci性xing收shou獲huo細xi胞bao、產物、培養基的操作方式。
該方式的特點:(1) 操作簡單。培養周期短,染菌和細胞突變的風險小。反應器係統屬於封閉式,培養過程中與外部環境沒有物料交換,除了控製溫度、pH值和通氣外,不進行其他任何控製,因此操作簡單,容易掌握;(2)直觀的反應細胞生長代謝的過程。 由於培養期間細胞的生長代謝是在一個相對固定的營養環境,不添加任何營養成分,因此可直觀的反應細胞生長代謝的過程,是動物細胞工藝基礎條件或"小試"研究常用的手段;(3)可(ke)直(zhi)接(jie)放(fang)大(da)。由(you)於(yu)培(pei)養(yang)過(guo)程(cheng)工(gong)藝(yi)簡(jian)單(dan),對(dui)設(she)備(bei)和(he)控(kong)製(zhi)的(de)要(yao)求(qiu)較(jiao)低(di),設(she)備(bei)的(de)通(tong)用(yong)性(xing)強(qiang),反(fan)應(ying)器(qi)參(can)數(shu)的(de)放(fang)大(da)原(yuan)理(li)和(he)過(guo)程(cheng)控(kong)製(zhi),比(bi)較(jiao)其(qi)它(ta)培(pei)養(yang)係(xi)統(tong)較(jiao)易(yi)理(li)解(jie)和(he)掌(zhang)握(wo),在(zai)工(gong)業(ye)化(hua)生(sheng)產(chan)中(zhong)分(fen)批(pi)式(shi)操(cao)作(zuo)是(shi)傳(chuan)統(tong)的(de)、常用的方法,其工業反應器(Genetech)規模可達12000L。
分批培養過程中,細胞的生長分為五個階段:延滯期、對數生長期、減速期、平穩期和衰退期。分批培養的周期時間多在3~5天,細胞生長動力學表現為細胞先經曆對數生長期(48~72h)細xi胞bao密mi度du達da到dao最zui高gao值zhi後hou,由you於yu營ying養yang物wu質zhi耗hao劫jie或huo代dai謝xie毒du副fu產chan物wu的de累lei積ji細xi胞bao生sheng長chang進jin入ru衰shuai退tui期qi進jin而er死si亡wang,表biao現xian出chu典dian型xing的de生sheng長chang周zhou期qi。收shou獲huo產chan物wu通tong常chang是shi在zai細xi胞bao快kuai要yao死si亡wang前qian或huo已yi經jing死si亡wang後hou進jin行xing。
經(jing)過(guo)改(gai)進(jin)的(de)攪(jiao)拌(ban)式(shi)生(sheng)物(wu)反(fan)應(ying)器(qi),目(mu)前(qian)仍(reng)是(shi)大(da)規(gui)模(mo)培(pei)養(yang)動(dong)物(wu)細(xi)胞(bao)用(yong)以(yi)生(sheng)產(chan)各(ge)種(zhong)藥(yao)物(wu)的(de)主(zhu)要(yao)設(she)備(bei),也(ye)是(shi)早(zao)期(qi)用(yong)以(yi)生(sheng)產(chan)單(dan)抗(kang)的(de)主(zhu)要(yao)途(tu)徑(jing)。在(zai)用(yong)攪(jiao)拌(ban)式(shi)生(sheng)物(wu)反(fan)應(ying)器(qi)分(fen)批(pi)式(shi)培(pei)養(yang)單(dan)抗(kang)中(zhong),最(zui)多(duo)采(cai)用(yong)的(de)是(shi)微(wei)囊(nang)或(huo)巨(ju)載(zai)體(ti)培(pei)養(yang)。與(yu)一(yi)般(ban)的(de)懸(xuan)浮(fu)培(pei)養(yang)比(bi)較(jiao),雜(za)交(jiao)瘤(liu)細(xi)胞(bao)依(yi)托(tuo)微(wei)囊(nang)化(hua)或(huo)巨(ju)載(zai)體(ti)後(hou),相(xiang)對(dui)固(gu)定(ding)化(hua),降(jiang)低(di)了(le)攪(jiao)拌(ban)培(pei)養(yang)時(shi)對(dui)細(xi)胞(bao)的(de)剪(jian)切(qie)力(li),提(ti)高(gao)了(le)細(xi)胞(bao)的(de)密(mi)度(du)和(he)穩(wen)定(ding)性(xing)及(ji)生(sheng)產(chan)率(lv)。在(zai)1986年以前,采用此種方式培養的雜交瘤細胞就有100多種。
2. 流加式操作(fed-batch culture)
流(liu)加(jia)式(shi)操(cao)作(zuo)是(shi)在(zai)批(pi)式(shi)操(cao)作(zuo)的(de)基(ji)礎(chu)上(shang),采(cai)用(yong)機(ji)械(xie)攪(jiao)拌(ban)式(shi)生(sheng)物(wu)反(fan)應(ying)器(qi)係(xi)統(tong),懸(xuan)浮(fu)培(pei)養(yang)細(xi)胞(bao)或(huo)以(yi)懸(xuan)浮(fu)微(wei)載(zai)體(ti)培(pei)養(yang)貼(tie)壁(bi)細(xi)胞(bao),細(xi)胞(bao)初(chu)始(shi)接(jie)種(zhong)的(de)培(pei)養(yang)基(ji)體(ti)積(ji)一(yi)般(ban)為(wei)終(zhong)體(ti)積(ji)的(de)1/2 ~1/3,在(zai)培(pei)養(yang)過(guo)程(cheng)中(zhong)根(gen)據(ju)細(xi)胞(bao)對(dui)營(ying)養(yang)物(wu)質(zhi)的(de)不(bu)斷(duan)消(xiao)耗(hao)和(he)需(xu)求(qiu),流(liu)加(jia)濃(nong)縮(suo)的(de)營(ying)養(yang)物(wu)或(huo)培(pei)養(yang)基(ji),從(cong)而(er)使(shi)細(xi)胞(bao)持(chi)續(xu)生(sheng)長(chang)至(zhi)較(jiao)高(gao)的(de)密(mi)度(du),目(mu)標(biao)產(chan)品(pin)達(da)到(dao)較(jiao)高(gao)的(de)水(shui)平(ping),整(zheng)個(ge)培(pei)養(yang)過(guo)程(cheng)沒(mei)有(you)流(liu)出(chu)或(huo)回(hui)收(shou),通(tong)常(chang)在(zai)細(xi)胞(bao)進(jin)入(ru)衰(shuai)亡(wang)期(qi)或(huo)衰(shuai)亡(wang)期(qi)後(hou)進(jin)行(xing)終(zhong)止(zhi)回(hui)收(shou)整(zheng)個(ge)反(fan)應(ying)體(ti)係(xi),分(fen)離(li)細(xi)胞(bao)和(he)細(xi)胞(bao)碎(sui)片(pian),濃(nong)縮(suo)、純化目標蛋白。
流加培養主要有以下特點:
1) 流加培養根據細胞生長速率、營ying養yang物wu消xiao耗hao和he代dai謝xie產chan物wu抑yi製zhi情qing況kuang,流liu加jia濃nong縮suo的de營ying養yang培pei養yang基ji,流liu加jia的de速su率lv通tong常chang與yu消xiao耗hao的de速su率lv相xiang同tong,根gen據ju測ce得de的de底di物wu濃nong度du控kong製zhi相xiang應ying的de流liu加jia過guo程cheng,以yi保bao證zheng合he理li的de培pei養yang環huan境jing與yu較jiao低di的de代dai謝xie產chan物wu抑yi製zhi水shui平ping。
2) 培養過程以低稀釋率流加,細胞在培養係統中停留時間較長,總細胞密度較高,產物濃度較高。
3) 流liu加jia培pei養yang過guo程cheng須xu掌zhang握wo細xi胞bao生sheng長chang動dong力li學xue,能neng量liang代dai謝xie動dong力li學xue,研yan究jiu細xi胞bao環huan境jing變bian化hua時shi的de瞬shun間jian行xing為wei。流liu加jia培pei養yang細xi胞bao培pei養yang基ji的de設she計ji和he培pei養yang條tiao件jian與yu環huan境jing優you化hua,是shi整zheng個ge培pei養yang工gong藝yi中zhong的de主zhu要yao內nei容rong。
4) 在工業化生產,懸浮流加培養工藝參數的放大原理和過程控製,比較其它培養係統較易理解和掌握,可采用工藝參數的直接放大。
liujiapeiyanggongyishidangqiandongwuxibaopeiyanggongyizhongzhanyouzhuliuyoushidepeiyanggongyi,yeshijinnianlaidongwuxibaodaguimopeiyangyanjiuderedian。liujiapeiyanggongyizhongdeguanjianjishushijichupeiyangjiheliujianongsuodeyingyangpeiyangji。tongchangjinxingliujiadeshijianduozaizhishushengchanghouqi,xibaozaijinrushuaituiqizhiqian,tianjiagaonongdudeyingyangwuzhi。keyitianjiayici,yeketianjiaduoci,weilezhuiqiugenggaodexibaomiduwangwangxuyaotianjiayiciyishang,zhizhixibaomidubuzaitigao;可進行脈衝式添加,也可以降低的速率緩慢進行添加,但為了盡可能的維持相對穩定的營養物質環境,後者采用較多;tianjiadechengfenbijiaoduo,fanshicuxibaoshengchangdewuzhijunkeyijinxingtianjia。liujiadezongtiyuanzeshiweichixibaoshengchangxiangduiwendingdepeiyanghuanjing,yingyangchengfenjibuguoshengerchanshengdaliangdedaixiefuchanwuzaochengyingyangliyongxiaolvxiajiangerchengweiwuxiaodeliyong;也不缺乏導致細胞生長抑製或死亡。
流加工藝中的營養成分主要分為三大類:
1) putaotang。putaotangshixibaodegongnengwuzhihezhuyaodetanyuanwuzhi,ranerdangqinongdujiaogaoshihuichanshengdaliangdedaixiechanwurusuan,yinerxuyaojinxingqinongdukongzhi,yizugouweichixibaoshengchangerbuzhiyuchanshengdaliangdefuchanwudenongduweijia;
2) 穀gu氨an酰xian胺an。穀gu氨an酰xian胺an是shi細xi胞bao的de供gong能neng物wu質zhi和he主zhu要yao的de氮dan源yuan物wu質zhi,然ran而er當dang其qi濃nong度du較jiao高gao是shi會hui產chan生sheng大da量liang的de代dai謝xie產chan物wu氨an,因yin而er也ye需xu要yao進jin行xing其qi濃nong度du控kong製zhi,以yi足zu夠gou維wei持chi細xi胞bao生sheng長chang而er不bu至zhi於yu產chan生sheng大da量liang的de副fu產chan物wu的de濃nong度du為wei佳jia;
3) 氨基酸、維生素及其他。主要包括營養必需氨基酸、營養非必需氨基酸、一些特殊的氨基酸如羥脯氨酸、羧基穀氨酸和磷酸絲氨酸;此外還包括其他營養成分如膽堿、生sheng長chang刺ci激ji因yin子zi。添tian加jia的de氨an基ji酸suan形xing式shi多duo為wei左zuo旋xuan氨an基ji酸suan,因yin而er多duo以yi鹽yan或huo前qian體ti的de形xing式shi替ti代dai單dan分fen子zi氨an基ji酸suan,或huo者zhe添tian加jia四si肽tai或huo短duan肽tai的de形xing式shi。在zai進jin行xing添tian加jia時shi,不bu溶rong性xing氨an基ji酸suan如ru胱guang氨an酸suan、酪氨酸和色氨酸隻在中性pH值部分溶解,可采用泥漿的形式進行脈衝式添加;其他的可溶性氨基酸以溶液的形式用蠕動泵進行緩慢連續流加。
流加式操作分為兩種類型:單一補料分批式操作和反複補料分批式操作。
1) 單(dan)一(yi)補(bu)料(liao)分(fen)批(pi)式(shi)操(cao)作(zuo)是(shi)在(zai)培(pei)養(yang)開(kai)始(shi)時(shi)投(tou)入(ru)一(yi)定(ding)量(liang)的(de)基(ji)礎(chu)培(pei)養(yang)液(ye),培(pei)養(yang)到(dao)一(yi)定(ding)時(shi)期(qi),開(kai)始(shi)連(lian)續(xu)補(bu)加(jia)濃(nong)縮(suo)營(ying)養(yang)物(wu)質(zhi),直(zhi)到(dao)培(pei)養(yang)液(ye)體(ti)積(ji)達(da)到(dao)生(sheng)物(wu)反(fan)應(ying)器(qi)的(de)最(zui)大(da)操(cao)作(zuo)容(rong)積(ji),停(ting)止(zhi)補(bu)加(jia),最(zui)後(hou)將(jiang)細(xi)胞(bao)培(pei)養(yang)液(ye)一(yi)次(ci)全(quan)部(bu)放(fang)出(chu)。該(gai)操(cao)作(zuo)方(fang)式(shi)受(shou)到(dao)反(fan)應(ying)器(qi)操(cao)作(zuo)容(rong)積(ji)的(de)限(xian)製(zhi),培(pei)養(yang)周(zhou)期(qi)隻(zhi)能(neng)控(kong)製(zhi)在(zai)較(jiao)短(duan)的(de)時(shi)間(jian)內(nei)。
2) 反(fan)複(fu)補(bu)料(liao)分(fen)批(pi)式(shi)操(cao)作(zuo)是(shi)在(zai)單(dan)一(yi)補(bu)料(liao)分(fen)批(pi)式(shi)操(cao)作(zuo)的(de)基(ji)礎(chu)上(shang),每(mei)個(ge)一(yi)定(ding)時(shi)間(jian)按(an)一(yi)定(ding)比(bi)例(li)放(fang)出(chu)一(yi)部(bu)分(fen)培(pei)養(yang)液(ye),是(shi)培(pei)養(yang)液(ye)體(ti)積(ji)始(shi)終(zhong)不(bu)超(chao)過(guo)反(fan)應(ying)器(qi)的(de)最(zui)大(da)操(cao)作(zuo)容(rong)積(ji),從(cong)而(er)在(zai)理(li)論(lun)上(shang)可(ke)以(yi)延(yan)長(chang)培(pei)養(yang)周(zhou)期(qi),直(zhi)至(zhi)培(pei)養(yang)效(xiao)率(lv)下(xia)降(jiang),才(cai)將(jiang)培(pei)養(yang)液(ye)全(quan)部(bu)放(fang)出(chu)。
3.半連續式操作(semi-continuous culture)、重複批式操作(repeated batch culture)、換液操作(medium change culture) 半(ban)連(lian)續(xu)式(shi)操(cao)作(zuo)又(you)稱(cheng)為(wei)重(zhong)複(fu)分(fen)批(pi)式(shi)操(cao)作(zuo)或(huo)換(huan)液(ye)操(cao)作(zuo)。采(cai)用(yong)機(ji)械(xie)攪(jiao)拌(ban)式(shi)生(sheng)物(wu)反(fan)應(ying)器(qi)係(xi)統(tong),懸(xuan)浮(fu)培(pei)養(yang)形(xing)式(shi)。在(zai)細(xi)胞(bao)增(zeng)長(chang)和(he)產(chan)物(wu)形(xing)成(cheng)過(guo)程(cheng)中(zhong),每(mei)間(jian)隔(ge)一(yi)段(duan)時(shi)間(jian),從(cong)中(zhong)取(qu)出(chu)部(bu)分(fen)培(pei)養(yang)物(wu),再(zai)用(yong)新(xin)的(de)培(pei)養(yang)液(ye)補(bu)足(zu)到(dao)原(yuan)有(you)體(ti)積(ji),使(shi)反(fan)應(ying)器(qi)內(nei)的(de)總(zong)體(ti)積(ji)不(bu)變(bian)。
這(zhe)種(zhong)類(lei)型(xing)的(de)操(cao)作(zuo)是(shi)將(jiang)細(xi)胞(bao)接(jie)種(zhong)一(yi)定(ding)體(ti)積(ji)的(de)培(pei)養(yang)基(ji),讓(rang)其(qi)生(sheng)長(chang)至(zhi)一(yi)定(ding)的(de)密(mi)度(du),在(zai)細(xi)胞(bao)生(sheng)長(chang)至(zhi)最(zui)大(da)密(mi)度(du)之(zhi)前(qian),用(yong)新(xin)鮮(xian)的(de)培(pei)養(yang)基(ji)稀(xi)釋(shi)培(pei)養(yang)物(wu),每(mei)次(ci)稀(xi)釋(shi)反(fan)應(ying)器(qi)培(pei)養(yang)體(ti)積(ji)的(de)1/2~3/4,以yi維wei持chi細xi胞bao的de指zhi數shu生sheng長chang狀zhuang態tai,隨sui著zhe稀xi釋shi率lv的de增zeng加jia培pei養yang體ti積ji逐zhu步bu增zeng加jia。或huo者zhe在zai細xi胞bao增zeng長chang和he產chan物wu形xing成cheng過guo程cheng中zhong,每mei隔ge一yi定ding時shi間jian,定ding期qi取qu出chu部bu分fen培pei養yang物wu,或huo是shi條tiao件jian培pei養yang基ji,或huo是shi連lian同tong細xi胞bao、載(zai)體(ti)一(yi)起(qi)取(qu)出(chu),然(ran)後(hou)補(bu)加(jia)細(xi)胞(bao)或(huo)載(zai)體(ti),或(huo)是(shi)新(xin)鮮(xian)的(de)培(pei)養(yang)基(ji)繼(ji)續(xu)進(jin)行(xing)培(pei)養(yang)的(de)一(yi)種(zhong)操(cao)作(zuo)模(mo)式(shi)。剩(sheng)餘(yu)的(de)培(pei)養(yang)物(wu)可(ke)作(zuo)為(wei)種(zhong)子(zi),繼(ji)續(xu)培(pei)養(yang),從(cong)而(er)可(ke)維(wei)持(chi)反(fan)複(fu)培(pei)養(yang),而(er)無(wu)需(xu)反(fan)應(ying)器(qi)的(de)清(qing)洗(xi)、消毒等一係列複雜的操作。在半連續式操作中由於細胞適應了生物反應器的培養環境和相當高的接種量,經過幾次的稀釋、換液培養過程,細胞密度常常會提高。
半連續式操作的特點為:1)培養物的體積逐步增加;2)可進行多次收獲;3)細胞可持續指數生長,並可保持產物和細胞在一較高的濃度水平,培養過程可延續到很長時間。
該(gai)操(cao)作(zuo)方(fang)式(shi)的(de)優(you)點(dian)是(shi)操(cao)作(zuo)簡(jian)便(bian),生(sheng)產(chan)效(xiao)率(lv)高(gao),可(ke)長(chang)時(shi)期(qi)進(jin)行(xing)生(sheng)產(chan),反(fan)複(fu)收(shou)獲(huo)產(chan)品(pin),可(ke)使(shi)細(xi)胞(bao)密(mi)度(du)和(he)產(chan)品(pin)產(chan)量(liang)一(yi)直(zhi)保(bao)持(chi)在(zai)較(jiao)高(gao)的(de)水(shui)平(ping)。在(zai)動(dong)物(wu)細(xi)胞(bao)培(pei)養(yang)和(he)藥(yao)品(pin)生(sheng)產(chan)中(zhong)被(bei)廣(guang)泛(fan)應(ying)用(yong)。
4. 連續式操作(continuous culture)
lianxushicaozuoshiyizhongchangjiandexuanfupeiyangmoshi,caiyongjixiejiaobanshishengwufanyingqixitong。gaimoshishijiangxibaojiezhongyuyidingtijidepeiyangjihou,weilefangzhishuaituiqidechuxian,zaixibaodazuidamiduzhiqian,yiyidingsuduxiangshengwufanyingqilianxutianjiaxinxianpeiyangji;yucitongshi,hanyouxibaodepeiyangwuyixiangtongdesudulianxucongfanyingqiliuchu,yibaochipeiyangtijidehengding。lilunshangjiang,gaiguochengkewuxianyanxuxiaqu。lianxupeiyangdezuidayoudianshifanyingqidepeiyangzhuangtaikeyidadaohengding,xibaozaiwendingzhuangtaixiashengchang。wendingzhuangtaikeyouxiaodeyanchangfenpipeiyangzhongdeduishushengchangqi。zaiwendingzhuangtaixiaxibaosuochudehuanjingtiaojianruyingyangwuzhinongdu、產物濃度、pH值(zhi)可(ke)保(bao)持(chi)恒(heng)定(ding),細(xi)胞(bao)濃(nong)度(du)以(yi)及(ji)細(xi)胞(bao)比(bi)生(sheng)長(chang)速(su)率(lv)可(ke)維(wei)持(chi)不(bu)變(bian)。細(xi)胞(bao)很(hen)少(shao)受(shou)到(dao)培(pei)養(yang)環(huan)境(jing)變(bian)化(hua)帶(dai)來(lai)的(de)生(sheng)理(li)影(ying)響(xiang),特(te)別(bie)是(shi)生(sheng)物(wu)反(fan)應(ying)器(qi)的(de)主(zhu)要(yao)營(ying)養(yang)物(wu)質(zhi)葡(pu)萄(tao)糖(tang)和(he)穀(gu)氨(an)酰(xian)胺(an),維(wei)持(chi)在(zai)一(yi)個(ge)較(jiao)低(di)的(de)水(shui)平(ping),從(cong)而(er)使(shi)他(ta)們(men)的(de)利(li)用(yong)效(xiao)率(lv)提(ti)高(gao),有(you)害(hai)產(chan)物(wu)積(ji)累(lei)有(you)所(suo)減(jian)少(shao)。然(ran)而(er)在(zai)高(gao)的(de)稀(xi)釋(shi)率(lv)下(xia),雖(sui)然(ran)死(si)細(xi)胞(bao)和(he)細(xi)胞(bao)碎(sui)片(pian)及(ji)時(shi)清(qing)除(chu),細(xi)胞(bao)活(huo)性(xing)高(gao)最(zui)終(zhong)細(xi)胞(bao)密(mi)度(du)得(de)到(dao)提(ti)高(gao);可是產物卻不斷在稀釋,因而產物濃度並為提高;尤其是細胞和產物不斷的稀釋,營養物質利用率、細胞增長速率和產物生產速率低下。此外,連續式操作還有一些不足,如:1)由於是開放式操作,加上培養周期較長,容易造成汙染;2)在長周期的連續培養中,細胞的生長特性以及分泌產物容易變異;3)對設備、儀器的控製技術要求較高。
連續式操作使用的反應器多數是攪拌式生物反應器,也可以是管式反應器。
連續式操作的特點為:1)細胞維持持續指數增長;2)產物體積不斷增長;3)可控製衰退期與下降期。
5. 灌流式操作(perfusion culture)
guanliushicaozuoshibaxibaohepeiyangjiyiqijiarufanyingqihou,zaixibaozengchanghechanwuxingchengguochengzhong,buduandijiangbufentiaojianpeiyangjiquchu,tongshiyoulianxubuduandiguanzhuxindepeiyangji。tayubanlianxushicaozuodebutongzhichuzaiyuquchubufentiaojianpeiyangjishi,juedabufenxibaojunbaoliuzaifanyingqinei,erbanlianxupeiyangzaiqupeiyangwushitongshiyequchulebufenxibao。
灌(guan)流(liu)式(shi)操(cao)作(zuo)常(chang)使(shi)用(yong)的(de)生(sheng)物(wu)反(fan)應(ying)器(qi)主(zhu)要(yao)有(you)兩(liang)種(zhong)形(xing)式(shi)。一(yi)種(zhong)是(shi)用(yong)攪(jiao)拌(ban)式(shi)生(sheng)物(wu)反(fan)應(ying)器(qi)懸(xuan)浮(fu)培(pei)養(yang)細(xi)胞(bao),這(zhe)種(zhong)反(fan)應(ying)器(qi)必(bi)須(xu)具(ju)有(you)細(xi)胞(bao)截(jie)流(liu)裝(zhuang)置(zhi),細(xi)胞(bao)截(jie)留(liu)係(xi)統(tong)開(kai)始(shi)多(duo)采(cai)用(yong)微(wei)孔(kong)膜(mo)過(guo)濾(lv)或(huo)旋(xuan)轉(zhuan)膜(mo)係(xi)統(tong),最(zui)近(jin)開(kai)發(fa)的(de)有(you)各(ge)種(zhong)形(xing)式(shi)的(de)沉(chen)降(jiang)係(xi)統(tong)或(huo)透(tou)析(xi)係(xi)統(tong)。
zhongkongxianweishengwufanyingqishilianxuguanliucaozuochangyongdeyizhong。tacaiyongdezhongkongxianweibantoumo,touguoxiaofenziliangdechanwuhediwu,jieliuxibaohefenziliangjiaodadechanwu,zailianxuguanliuguochengzhongjiangjuedabufenxibaojieliuzaifanyingqinei;近年來中空纖維生物反應器被廣泛應用於產物分泌性動物細胞的生產,主要用於培養雜交瘤細胞生產單克隆抗體。
另ling外wai一yi種zhong形xing式shi是shi固gu定ding床chuang或huo流liu化hua床chuang生sheng物wu反fan應ying器qi,固gu定ding床chuang是shi在zai反fan應ying器qi中zhong裝zhuang配pei固gu定ding的de籃lan筐kuang,中zhong間jian裝zhuang填tian聚ju脂zhi纖xian維wei載zai體ti,細xi胞bao可ke附fu著zhe在zai載zai體ti上shang生sheng長chang,也ye可ke固gu定ding在zai載zai體ti纖xian維wei之zhi間jian,靠kao上shang攪jiao拌ban中zhong產chan生sheng的de負fu壓ya,迫po使shi培pei養yang基ji不bu斷duan流liu經jing填tian料liao,有you利li於yu營ying養yang成cheng分fen和he氧yang的de傳chuan遞di,這zhe種zhong形xing式shi的de灌guan流liu速su度du較jiao大da,細xi胞bao在zai載zai體ti中zhong高gao密mi度du生sheng長chang。流liu化hua床chuang生sheng物wu反fan應ying器qi是shi通tong過guo流liu體ti的de上shang升sheng運yun動dong使shi固gu體ti顆ke粒li維wei持chi在zai懸xuan浮fu狀zhuang態tai進jin行xing反fan應ying,適shi合he於yu固gu定ding化hua細xi胞bao的de培pei養yang。
灌流式操作的共同優點是:①細胞截流係統可使細胞或酶保留在反應器內,維持較高的細胞密度,一般可達107-109/ml,從而較大的提高了產品的產量;②連續灌流係統,使細胞穩定的處在較好的的營養環境中,有害代謝廢物濃度積累較低;③反應速率容易控製,培養周期較長,可提高生產率,目標產品回收率高;④產(chan)品(pin)在(zai)罐(guan)內(nei)停(ting)留(liu)時(shi)間(jian)短(duan),可(ke)及(ji)時(shi)回(hui)收(shou)到(dao)低(di)溫(wen)下(xia)保(bao)存(cun),有(you)利(li)於(yu)保(bao)持(chi)產(chan)品(pin)的(de)活(huo)性(xing)。連(lian)續(xu)灌(guan)注(zhu)培(pei)養(yang)法(fa)是(shi)近(jin)年(nian)來(lai)用(yong)於(yu)哺(bu)乳(ru)動(dong)物(wu)細(xi)胞(bao)培(pei)養(yang)生(sheng)產(chan)分(fen)泌(mi)型(xing)重(zhong)組(zu)治(zhi)療(liao)性(xing)藥(yao)物(wu)和(he)嵌(qian)合(he)抗(kang)體(ti),以(yi)及(ji)人(ren)源(yuan)化(hua)抗(kang)體(ti)等(deng)基(ji)因(yin)工(gong)程(cheng)抗(kang)體(ti)較(jiao)為(wei)推(tui)崇(chong)的(de)一(yi)種(zhong)操(cao)作(zuo)方(fang)式(shi)。應(ying)用(yong)連(lian)續(xu)灌(guan)流(liu)工(gong)藝(yi)的(de)公(gong)司(si)有(you)Genzyme, Genetic Institute, Bayer公司等
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