CN105477650A - 使用纳米纤维过滤介质从流体样品除去微生物 - Google Patents
使用纳米纤维过滤介质从流体样品除去微生物 Download PDFInfo
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Abstract
从液体样品除去微生物的方法和包含纳米纤维的液体过滤介质,其同时呈现出高液体渗透率和高微生物截留率。使液体通过多孔的包含纳米纤维的过滤介质,从所述液体除去微生物如细菌,特别是缺陷短波单胞菌,所述过滤介质的缺陷短波单胞菌LRV大于约9,所述纳米纤维的直径为约10nm-约1,000nm。除去微生物如细菌和支原体的另一方法包括使液体通过多孔的包含纳米纤维的过滤介质,所述过滤介质的微生物LRV大于约8,并且所述纳米纤维的直径为约10nm-约1,000nm。所述过滤介质可以是纤维性电纺聚合物纳米纤维液体过滤介质垫的形式。
Description
相关申请的交叉引用
本申请是2010年3月19日递交的国际申请号为PCT/US2010/000826,中国国家申请号为201080017401.8,名称为“使用纳米纤维过滤介质从流体样品除去微生物”的申请的分案申请。本申请的母案申请要求于2009年3月19日提交的美国临时专利申请61/210,468的权利。
技术领域
本发明一般地涉及过滤介质。在某些实施方案中,本发明提供多孔电纺纳米纤维液体过滤垫,以及使用所述过滤垫从被过滤的液体截留微生物的方法。
背景技术
用于液体过滤的过滤器通常可归类为纤维性无纺介质过滤器或多孔膜过滤器。
纤维性无纺液体过滤介质包括但不限于由纺粘的,熔喷的或水刺(spunlace)的连续纤维制得的无纺介质;由粗梳的短纤维等制得的水刺(hydroentangle)无纺介质;或者这些种类的一些组合。典型地,用于液体过滤的纤维性无纺过滤介质过滤器的孔径通常大于约1微米(μm)。
多孔膜液体过滤介质在没有支持体下使用,或者与多孔基材或支持体一起使用。多孔过滤膜的孔径小于纤维性无纺介质,典型地,其孔径小于约1μm。多孔膜液体过滤膜可用于:(a)微过滤,其中从液体中被过滤的颗粒物典型地为约0.1μm-约10μm;(b)超滤,其中从液体中被过滤的颗粒物典型地为约5nm-约0.1μm;和(c)反渗透,其中从液体中被颗粒的颗粒物质典型地为约-约1nm。
纤维性无纺介质和多孔膜都适合用于微过滤。在工业中,微过滤被广泛地认为是从流体流除去微生物如细菌的可靠的易于改变规模的有益方法,并且是制药和生物制药中必不可少的部分。在生物制药业中特别重要,在生物制药过程中多处使用微过滤。
但是,为了通过使用纤维性无纺介质的微过滤达到相当于孔径小于约1μm的颗粒截留,需要增大过滤器中纤维材料的层数以增大无纺介质的深度。增大无纺介质中的纤维层数会同时产生期望的和不令人期望的结果。增大纤维层数,由于使污染物颗粒必须经过其免除被过滤介质捕获的缺陷途径的迂曲度增大,并使过滤介质的保持污染物的能力增大,由此产生期望的结果。但是,增大无纺介质中的纤维层数不令人期望地使在使用时经过该介质的压降或压差增大,这意味着增大用户使用过滤器的能量并且缩短过滤器的寿命。
用于微过滤的多孔膜过滤器,不同于纤维性无纺介质,提供良好的颗粒截留率、压降和通量的组合,但是往往成本过高,并且典型地不提供在整个压降范围内的良好的保持污染物能力,因此限制多孔膜过滤器的寿命。
液体微过滤膜的两个最期望的特征是高渗透率和可靠的截留率。当然,存在这两个参数之间的平衡,并且对于相同类型的膜,过去通过牺牲膜的渗透率来获得更大的截留率。制备膜的常规方法的固有局限是妨碍膜超过孔隙率的某阈值,因此在特定孔径下可达到的渗透率的大小受到限制。
过滤膜截留微生物的定量量度通常表达为对数下降值(LogReductionValue)或LRV。LRV是挑战试验溶液(challengesolution)中的颗粒浓度比过滤器流出液中的颗粒浓度的比值的对数:
LRV=Log{[CFU]挑战/[CFU]流出液}
在过滤器在测试条件下截留全部微生物的情况中,通常记录的LRV大于单种微生物通过过滤器时所得的值。例如,在4.77*107CFU/cm2的进行挑战试验性颗粒浓度下,最大可测得的LRV为8.22。当没有颗粒穿过过滤器时,LRV记录为大于8.22。
膜的孔径值是已成功通过相关的标准细菌进行挑战测试的膜的标识。最常见的孔径值为0.22μm,这评定为通过测定应用于液体过滤的膜过滤器的细菌截留的标准测试方法(StandardTestMethodforDeterminingBacterialRetentionOfMembraneFiltersUtilizedForLiquidFiltration,ASTMF838-83测试)的膜,可验证在被>107CFU/cm2缺陷短波单胞菌(Brevundimonasdiminuta)进行挑战试验后,所述膜产生无菌流出液。
缺陷短波单胞菌(ATTC#19146),以前称为缺陷假单胞菌(Pseudomonasdiminuta),是需氧革兰氏阴性细菌(杆菌)。由于其小尺寸,缺陷短波单胞菌(B.diminuta)是验证用于灭菌膜过滤器等的标准微生物。但是,虽然缺陷短波单胞菌是大多数病原菌的代表,已证明,对于称为支原体的一类微生物而言,缺陷短波单胞菌是差的模式。虽然缺陷短波单胞菌是大多数病原菌的代表,但已证明对于称为支原体的一类微生物而言是差的模式。
支原体是可感染细胞培养物并且可大幅有害地影响生物制药的微生物。真核细胞培养物等被支原体污染也是常见问题,产生不可靠的实验结果和可能不安全的生物产品。对于从事生物学和药学产品的开发和生产的生产者而言,这是严重的问题。用于细胞培养的介质的富含营养的环境可导致支原体繁殖,从而降低细胞增长并且损失培养物。不同于被可在感染后短期内依据可见影响检测到的细菌类的污染,这些细菌诸如致细胞病变性、pH变化、异常生长或呈现混浊的介质之类,由支原体所致的污染可能在没有明显症状的情况下未被检测到(Razin,S.1997.ComparativegenomicsofMycoplasmas.WienKlinWochenschr109:551-6.JungH.WangSY,YangIW,HsuchDW,YangWJ,WangTH,Wang:HS.(2003)DetectionandtreatmentofMycoplasmacontaminationinculturedcells.ChangGungMedJ.26:250-8.WisherM.(2002)Biosafetyandproductreleasetestingissuesrelevanttoreplication-competentoncolyticviruses,Review.CancerGeneTher.9:1056-61)。
孔径值为0.1μm的膜表明已验证除去支原体的膜(参见Roche,K.L.:Levy,R.V.,MethodstoValidateMicroporousMembranesfortheRemovalofMycoplasma,BioPharm1992,5,(3),22-33)。
例如,孔径值为0.1um的膜可用来过滤被供至在生物反应器内存活和生长的细胞的培养基、营养物和细胞培养流体。现有的膜对于拉氏无胆甾原体(A.Laidawii,支原体的测试微生物)具有特定的对数下降值(LRV)。虽然通常认为LRV>8足以断言"完全"截留支原体,但是由于渗透率较大并且通过量较高,LRV较小的过滤器反而常用于液体过滤中。
转让给MilliporeCorporation并且题为SERUM-FREEGROWTHMEDIUMFORACHOLEPLASMALAlDLAWIIANDMETHODSFORRETENTIONTESTINGSTERILIZINGGRADEFILTERS的WO/2009/032040,通过援引完全并入本文,教导在被≥1x109cfu/mL的拉氏无胆甾原体(Acholeplasmalaidlawii,A.laidlawii;ATCC23206)进行挑战试验后,产生无菌流出液,可验证过滤介质完全截留支原体。
例如,孔径值为0.1μm的两种膜VV和ExpressSHR(各可从MilliporeCorporation,Billerica,MA,USA获得)的支原体LRV分别为4和6。虽然也可从MilliporeCorporation获得的MV声称完全截留支原体(LRV>8),但是在培养基过滤时,它与VV和ExpressSHR相比具有较低的渗透率和容量。
利用包括熔喷、静电纺织和电吹在内的各种方法,合成聚合物已被制成直径非常小的纤维网,即,数微米或小于1μm的量级。已表明此类网有效地用作液体屏蔽材料和过滤器。通常它们与更强的片材一起形成复合材料,其中该更强的片材提供满足最终过滤器产品所需的强度。
授予Schaefer等人的美国专利公布2004/0038014教导用于过滤污染物的无纺过滤垫,其包含一层或多层通过静电纺织形成的细聚合物微纤维和纳米纤维的厚聚集物。静电纺织法利用电纺装置,其包括:储液槽(其中装有形成细纤维的聚合物溶液)、泵,以及喷射装置(其从储液槽获得聚合物溶液)。在静电场中,聚合物溶液的液滴被静电场加速,向位于格栅上的收集介质基材运动。利用适合的静电高压电源,在喷射器和格栅之间保持高电压的静电电位,并且所述收集基材介于其间。
"电吹"法公开于世界专利公布WO03/080905中,通过援引全文并入本文。将包含聚合物和溶剂的聚合物溶液流从储藏槽供至喷丝头内的一系列纺丝口,向其施加高电压并通过其喷出聚合物溶液。同时,任选地被加热的压缩空气从位于纺丝口的侧面或***的空气喷嘴喷出。空气通常被向下引导,并且作为吹气流包围并推进新喷射出的聚合物溶液并且促进形成纤维网,其被收集在位于真空室上方的接地的多孔收集带上。该电吹法能够在相对短期内形成基重超过约1gsm,甚至高达约40gsm或更高的商品尺寸和数量的纳米网。
授予Bates等人的美国专利公布2007/0075015教导用于过滤液体中的颗粒物质的液体过滤介质,其包含任选地置于粗布(scrim)层上的至少一层平均直径小于1,000纳米的纳米纤维。该过滤介质在相对高水平的紧密度(solidity)下流速为至少0.055L/min/cm2。该介质在压差增高2psi(14kPa)至15psi(100kPa)时流速没有明显降低。
授予Xu的美国专利公布2007/0018361教导通过反应型电纺制备纳米纤维,其中电纺法与在线反应器(其中进行化学或光化学反应)联用。Xu中教导的方法利用电纺能够由交联的聚合物及其它材料制备纳米纤维。
授予Chen的美国专利公布2009/0026137教导用包含与微孔膜相邻并任选地粘合的纳米网的复合介质制备液体过滤器。该膜的特征是在评定的粒径下LRV值为3.7,并且该纳米网在所述膜的评定粒径下具有大于0.1的分级过滤效率。该纳米网在所述效率下还具有大于0.0002的厚度效率比。该纳米网用来赋予该膜深度过滤。
期望具有可靠的电纺纳米纤维液体过滤介质,当从通过该过滤介质的液体除去微生物时,其微生物LRV大于约8,适合完全截留微生物如细菌,特别是支原体,和/或缺陷短波单胞菌;LRV大于约9,适合完全截留缺陷短波单胞菌,并且同时达到高渗透率和高通过量。
此外,该多孔电纺纳米纤维过滤介质易于改变尺寸,可适应于数毫升至数千升的样品流体的处理体积,并且能够与各种过滤方法和装置一起使用。本发明涉及这些,以及其它目的和实施方案。
发明内容
本发明涉及使液体通过多孔的电纺纳米纤维液体过滤介质,从而从该液体中除去微生物的方法。所述电纺纳米纤维液体过滤介质可被或不被置于多孔支持体或基材上使用。所述电纺纳米纤维液体过滤介质可被制成各种形状、尺寸、厚度和密度,例如,多孔的、聚合物纳米纤维垫。
在另一实施方案中,本发明涉及多孔电纺纳米纤维液体过滤介质,其缺陷短波单胞菌LRV大于约9,并且所述纳米纤维的平均纤维直径为约10nm-约1000nm。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其缺陷短波单胞菌LRV大于约9,并且所述过滤介质的孔隙率为约80%-约95%。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其支原体LRV大于约4,并且其在10psi压差下的液体渗透率大于约3,000LMH.(升每平方米每小时)。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其支原体LRV大于约8,并且其在10psi压差下的液体渗透率大于约3,000LMH。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其缺陷短波单胞菌LRV大于约9,并且被制成为厚度约1μm-约500μm的纤维多孔垫。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其缺陷短波单胞菌LRV大于约9,并且其在10psi压差下的液体渗透率大于约10,000LMH。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其支原体LRV大于约8,并且所述纳米纤维的平均纤维直径为约10nm-约1,000nm。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其支原体LRV大于约8,并且其在10psi压差下的液体渗透率大于约3,000LMH。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其支原体LRV大于约8,并且所述过滤介质的孔隙率为约80%-约95%。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其支原体LRV大于约8,并且被制成厚度约1μm-约500μm的纤维多孔垫。
在另一实施方案中,本发明涉及多孔的电纺纳米纤维液体过滤介质,其支原体LRV大于约8,并且其在10psi压差下的液体渗透率大于约10,000LMH。
在另一实施方案中,本发明涉及方法:通过使用电纺装置,由聚合物溶液中的一种或多种电纺聚合物纳米纤维形成多孔过滤介质,并对该溶液施加大于约10kV的电位,并且收集电纺聚合物纤维作为无纺垫。
在另一实施方案中,本发明涉及复合多孔过滤装置,其包含微生物LRV大于约8的过滤介质,并且包含置于多孔支持体或多孔基材上的电纺聚合物纳米纤维垫。
在以下详细的说明书和权利要求书中说明本发明的其它特征和优点。对本领域技术人员而言显而易见,在不脱离本发明的精神和范围的情况下,可对本发明进行许多修改和改变。应理解以上概述及以下详述、权利要求书和附图仅是示例性和解释性,并且意在提供本教导的各种实施方案的解释。只是借助于实施例体现本文所述的具体实施方案,而绝不意在限制。
附图说明
包括在本申请中并且作为本申请组成部分的附图说明用于解释本发明的目前考虑的实施方案,其和说明书一起用来解释本发明的原理。
图1是本发明的一个实施方案所述的电纺纳米纤维的方法的示意图。聚合物溶液10,转鼓20,移动收集带30,接地电极35,高压电源40,由电场50制得的聚合物纤维,由聚合物纤维形成的纤维垫60。
图2是来自实施例1中列举说明的本发明的一个实施方案的尼龙纤维的横截面扫描电子显微镜照片。
图3是来自实施例1中列举说明的本发明的一个实施方案的尼龙纤维的正面扫描电子显微镜照片。
图4是来自实施例2中列举说明的本发明的另一实施方案的尼龙纤维的扫描电子显微镜照片。
图5是来自对比例1的对称商品膜MVPP的扫描电子显微镜照片。
图6是来自对比例2的对称商品膜GVPP的扫描电子显微镜照片。
具体实施方式
本文无论在上文或下文中引用的所有出版物、专利和专利申请通过援引特此整个并入,正如各篇出版物、专利或专利申请被特意地分别指出通过援引并入本文。
除非另外指出,就本说明书和权利要求书而言,本说明书和权利要求书中使用的表述组分的量、材料的百分比或比例、反应条件的所有数值及其它数值应理解为在所有情况中被术语“约”修饰。
因此,除非相反地指出,以下说明书和随附权利要求书中所述的数值参数是可根据本发明寻求获得的期望性质改变的近似值。绝非试图限制将等同原则应用于本权利要求书的范围,应至少根据记录的有效数字的数值并应用常规的舍入法解释各数值参数。
尽管表述本发明的宽范围的数值域和数值参数是近似值,但是尽可能准确地记录具体实施例中所述的数值。然而,任何数值固有地包含由它们各自的实验测定中存在的标准偏差必然产生的某些误差。此外,本文公开的所有范围应理解为涵盖其中所含的所有子范围。例如,"1-10"包括最小值1与最大值10(包含端点)之间的任何和全部子范围,即,具有等于或大于1的最小值和等于或小于10的最大值的任何和所有子范围,例如,5.5-10。
在进一步详述本发明之前,对多个术语进行定义。这些术语的使用不是限制本发明的范围,而是仅为了便于描述本发明。
在本文使用时,除非上下文明确地另外指出,单数形式"一个"、"一种"和"该/此/这"包括复数的所指物。
术语"纳米纤维"是指直径为数十纳米至数百纳米但通常小于一微米的纤维。
术语"过滤媒介"或"过滤介质"是指当携带微生物污染物的流体通过材料或材料聚集物(collectionofmaterial)时其中的微生物沉积在所述材料或材料聚集物之中或之上的材料或材料聚集物。
术语"通量"和"流速"可互换地用来指某个体积的流体通过特定面积的过滤介质的速度。
本发明的过滤介质包含多孔的电纺纳米纤维液体过滤垫。所述纳米纤维的平均纤维直径为约10nm-约1000nm。所述过滤介质的平均孔径为约0.1μm-约1μm。所述过滤介质的孔隙率为约80%-约95%。所述过滤介质的厚度为约1μm-约500μm,优选为约50μm-约200μm。所述过滤介质的液体渗透率大于约300LMH/psi。
适合用于本发明的纳米纤维中的聚合物包括热塑型和热固型聚合物。适合的聚合物包括但不限于:尼龙、聚酰亚胺、脂肪族聚酰胺、芳香族聚酰胺、聚砜、纤维素、乙酸纤维素酯、聚醚砜、聚氨酯、聚(脲氨酯)(poly(ureaurethane))、聚苯并咪唑、聚醚酰亚胺、聚丙烯腈、聚(对苯二甲酸乙二醇酯)、聚丙烯、聚苯胺、聚(氧化乙烯)、聚(萘二甲酸乙二醇酯)、聚(对苯二甲酸丁二醇酯)、苯乙烯-丁二烯橡胶、聚苯乙烯、聚(氯乙烯)、聚(乙烯醇)、聚(偏二氟乙烯)、聚(乙烯基丁烯)、它们的共聚物、衍生化合物和混合物,及它们的组合。
制备过滤介质的电纺纳米纤维垫的方法公开于WO2005/024101;WO2006/131081和WO2008/106903中,均转让给捷克共和国Liberec的ElmarcoS.R.O.。
在本发明的一个实施方案中,所述过滤介质包含由单纳米纤维制成的垫,其中所述单纳米纤维是通过位于转鼓与收集器之间的移动收集装置单次经过此工序制得。应理解所述纤维网可通过一个或多个转鼓在同一移动收集装置上方同时运转制得。
在本发明的一个实施方案中,纤维垫通过从尼龙溶液沉积纳米纤维进行制备。在已蒸发或除去残余的溶剂后,在干燥基础上测定,所述纳米纤维垫的基重为约5g/m2-约15g/m2。
如图1中所示,移动收集装置30优选为位于转鼓20与收集器35之间的静电场内的移动收集带,其中收集由单纳米纤维制成的多孔垫。
在本发明的一个实施方案中,可将各种多孔的单层或多层的基材或支持体中的任一种安置于移动收集带上,以收集并与所述电纺纳米纤维垫介质结合,来形成复合过滤装置。
单层或多层的多孔的基材或支持体的实例包括但不限于纺粘无纺物(nonwoven)、熔喷无纺物、针刺无纺物、水刺无纺物(spunlacednonwoven)、湿法无纺物(wetlaidnonwoven)、树脂粘合无纺物、织造织物、针织织物、纸、及它们的组合。
在本发明的另一个实施方案中,本文所述的电纺纳米纤维垫介质可与多孔的基材或支持体粘合。粘合可通过现有技术中已知的方法(包括但不限于在加热的平滑压料辊之间热压延、超声焊合和气体粘合)完成。粘合增强所述介质的强度和耐压性,从而所述介质可耐受与被加工、被制成有用的过滤器和被用于过滤器中相关并取决于使用的粘合方法的力,调整物理性质,诸如厚度、密度和孔的形状和尺寸。
例如,热压延法可用来降低厚度,增大密度,降低电纺纳米纤维垫介质的孔隙率,以及缩小孔径。这继而降低在特定施加的压差下通过介质的流速。通常,超声焊合比热压延粘合更小面积的电纺纳米纤维垫介质,因此,对厚度、密度和孔径具有较小的影响。气体粘合通常对厚度、密度和孔径具有最小的影响,因此在期望保持较高流体流速的应用中可优选此粘合方法。
当采用热压延法时,必须注意,不要过度粘合所述电纺纳米纤维材料,以便所述纳米纤维熔解并不再截留它们作为单独纤维的结构。在极端情况中,过度粘合会导致所述纳米纤维完全熔解以至会形成膜。将使用的一个或两个压料辊加热至约环境温度(例如约25℃)至约300℃的温度。可在压力约0lb/in-约1000lb/in(178kg/cm)的压料辊之间压制所述纳米纤维垫和/或多孔支持体或基材。所述纳米纤维垫可以以至少约10ft/min(3m/min)的线速度压制。
可调整压延条件,例如,辊温度、辊隙压力(nippressure)和线速度达到期望的紧密度。通常施加较高的温度、压力和/或在升高的温度和/或压力下的滞留时间致使紧密度增高。
按照期望,在形成,塑形和制作所述电纺纳米纤维垫介质的整个方法中可任选地包括诸如拉伸、冷却、加热、烧结、退火、卷绕、拆卷等之类的其它机械步骤。
例如,按照期望,可在单步或多步步骤中拉伸本文所述的电纺纳米纤维垫介质。取决于用来拉伸所述电纺纳米纤维垫介质的拉伸方法,拉伸可调整包括厚度、密度以及垫中形成的孔的尺寸和形状在内的所述垫的物理性质。例如,若所述电纺纳米纤维垫在单方向上被拉伸(单轴拉伸),则可通过单步拉伸或一系列拉伸步骤进行拉伸直至获得期望的最终拉伸比。
相似地,若所述电纺纳米纤维垫介质在两个方向上被拉伸(双轴拉伸),则可通过单步双轴拉伸步骤或一系列双轴拉伸步骤进行拉伸直至获得期望的最终拉伸比。双轴拉伸还可通过一系列在一方向上的一步或多步单轴拉伸步骤和在另一方向上的一步或多步单轴拉伸步骤完成。可以任何顺序依次地进行双轴拉伸步骤(其中同时在两个方向上拉伸所述电纺纳米纤维垫)和单轴拉伸步骤。
拉伸所述垫的方法没有特别限制,并且可采用常规的拉幅、辊压或吹胀,或者这些中的两种或多种方法的组合。所述拉伸可以以单轴、双轴等方式进行。在双轴拉伸的情况中,纵向拉伸和横向拉伸可同时或相继地进行。
各种拉伸装置是本领域公知的,并且可用来完成本发明所述电纺垫的拉伸。单轴拉伸通常通过在两个辊之间拉伸进行,其中第二或下游辊旋转的圆周速度大于第一或上游辊。还可在标准拉幅机上进行单轴拉伸。
双轴拉伸可通过在拉幅机上在两个不同方向上同时拉伸完成。但是,更普遍地,通过首先在如上所述的两个差速旋转的辊之间单轴拉伸,然后使用拉幅机在不同的方向上单轴拉伸,或者通过使用拉幅机双轴拉伸,完成双轴拉伸。最普遍的双轴拉伸类型是其中两个拉伸方向彼此约成直角。在连续片材被拉伸的大多数情况中,一个拉伸方向至少大约平行于该片材的长轴(纵向),另一拉伸方向至少大约垂直于纵向并在该片材的面内(横向)。
在所述电纺纳米纤维垫已被单轴或双轴拉伸后,被拉伸的多孔的电纺纳米纤维垫可再被压延。为了形成与从拉伸装置出来的垫相比厚度减小的垫,可将被拉伸的电纺纳米纤维垫输送至一对配合作用的被加热的压延辊。通过调节这些压延辊施加的压力和温度,可按照期望控制最终电纺纳米纤维垫的孔径,由此能够调整平均孔径。
在拉伸之前,之时和/或之后,可通过多种方法中的任一种加热所述电纺纳米纤维垫。这些方法的实例包括:诸如由电加热的或燃气加热的红外线加热器提供的辐射加热,诸如由再循环热空气提供的对流加热,以及诸如通过与被加热的辊接触提供的传导加热。为了控温而测定的温度可能随着使用的装置和个人喜好而变。
通常,可以控制温度,从而所述电纺纳米纤维垫被大约均匀地拉伸,以至该被拉伸的垫的厚度差异(若有)在可接受的限度内,并且在那些限度之外的被拉伸的微孔电纺纳米纤维垫的量可接受地低。显然用于控制目的的温度可能接近于或不接近于所述电纺纳米纤维垫本身的那些温度,因为它们取决于使用的装置的性质、测温装置的位置以及被测温的物质或物体的特征。
可通过压延调整孔隙率。可获得约5%-约90%的孔隙率。虽然过滤介质通常采用单层结构,但有时配备彼此相邻的多于一层的过滤介质是有利的。改进颗粒截留率的分层式膜过滤器常用于病毒过滤,并且商业应用于NFP和Viresolve的Millipore产品生产线中。具有相同或不同的组成的分层式过滤介质还用来改进过滤器通过量。此类分层式过滤器的实例是Millipore的SHC和SHRP产品线。选择多层过滤产品的其它因素包括生产介质和装置的经济和方便性,灭菌和验证的容易性。本发明的纤维性过滤介质可采用单层或多层的结构。
测试方法
基重按照ASTMD-3776(特此通过援引并入)进行测定并以g/m2为单位记录。
孔隙率通过样品的基重(g/m2)除以聚合物密度(g/cm3),除以样品厚度(微米)乘以100,并从100中减去所得数进行计算,即,孔隙率=100-[基重/(密度×厚度)×100]。
纤维直径如下进行测定。以40,000倍放大率取得各纳米纤维层样品的10幅扫描电子显微镜(SEM)图像。从各SEM图像测得十根(10)清晰可分辨的纳米纤维的直径并记录。不包括缺陷(即,纳米纤维团、聚合物滴、纳米纤维交叉)。计算各样品的平均纤维直径。
厚度按照ASTMD1777-64(特此通过援引并入)进行测定并以微米记录。
平均流量起泡点的测定如下进行:按照ASTMDesignationE1294-89,“用自动液体孔率计检验薄膜过滤器的孔径特性的测试方法(StandardTestMethodforPoreSizeCharacteristicsofMembraneFiltersUsingAutomatedLiquidPorosimeter)”,利用来自ASTMDesignationF316的自动起泡点法,使用与来自PorousMaterials,Inc.(PMI),Ithaca,N.Y的商购装置原理相似的定制的毛细流式孔隙率测定仪进行测定。用异丙醇润湿直径47mm的各个样品(9.6cm2的可测面积)。将各样品置于支持物中,施加空气压差,并从样品中除去流体。利用提供的软件,使用在湿流量等于干流量(没有润湿溶剂的流量)的一半时的压差计算平均流量孔径。
流速(也称为通量)是流体通过特定面积的样品的速度,并通过使去离子水通过直径为35mm的过滤媒介样品进行测定。使用液压(水头压力)或气压(水上的空气压力)迫使水通过样品。
可使用诸如起泡点、液体-液体气孔测量法和特定粒径颗粒的挑战试验等常规的膜技术,测定电纺垫的有效孔径。已知纤维垫的有效孔径通常随着纤维直径增加并随着孔隙率降低。
起泡点测试提供测定有效孔径的方便方法。从以下方程计算:
其中P是起泡点压力,γ是探测流体(probefluid)的表面张力,r是孔半径,θ是液体-固体接触角。
膜生产商根据膜过滤器的截留特征确定商品膜过滤器的标称孔径值。
虽然已知无规无纺垫的孔径分布随着垫的厚度增大而变得更窄(参见Meltzer,T.H.,InFiltrationinthePharmaceuticalIndustry,MarcelDekker:NewYork,1987;p103),但之前未表明,在截留支原体的过滤器的至少100LMH/psi和截留缺陷短波单胞菌的过滤器的500LMH/psi的竞争性渗透率下,无纺垫的孔径分布是否可达到足够窄从而实现"完全截留细菌"(如上所述)。
通过用8.77*107菌落形成单位/平方厘米的膜(CFU/cm2)对膜进行挑战试验,从而测定支原体截留率。用50ml稀释的拉氏无胆甾原体对这些装置进行挑战试验,然后用50ml支原体缓冲液冲洗,总计100ml。然后通过0.22μm灭菌膜过滤全部100ml。然后,采用已公布的专利申请WO2009/032040中所述的方法。
缺陷短波单胞菌截留率按照ASTMF838-83进行测定。
本发明的以下实施例表明电纺纳米纤维垫可同时兼具高渗透率和高细菌截留率。
下文中在以下实施例中进一步详述本发明。通过以下意在列举说明本发明的实施例进一步说明本发明。
实施例
实施例1
采用WO2006/131081中公开的制备纳米纤维网的电纺法和装置,来制备以下实施例的纳米纤维层和垫。
通过电纺尼龙6聚合物溶液制得纳米纤维层。尼龙6由BASFCorp.,FlorhamPark,NJ,USA以商品名UltramidB24供应。使用乙酸和甲酸的溶剂混合物(重量比2:1)配制浓度为8%-16%的尼龙溶液。
在82kV下,并且溶液与接地电极的间距为155mm,电纺10重量%的尼龙溶液,持续45分钟。仅作为示例,使用上述标准Millipore方法,测试样品的拉氏无胆甾原体截留率。在下表I中,将代表性样品与最接近的膜MVPP比较。
结果示于下表I中。
表I
实施例2
按照实施例1中所述,制得一系列尼龙6电纺纤维垫。在82kV下,并且溶液与接地电极的间距为155mm,电纺13重量%的尼龙溶液,分别持续10和45分钟。分别对应于两种纺织时间,制得厚度为55微米和225微米的纤维垫。仅作为示例,测试这些样品的缺陷短波单胞菌截留率。应注意本文教导的截留支原体的电纺纤维垫用于完全截留缺陷短波单胞菌。
实施例3
按照实施例2中所述,制得另一系列的尼龙6电纺纤维垫。在82kV下,并且溶液与接地电极的间距为155mm,电纺16重量%的尼龙溶液,持续15分钟。仅作为示例,测试这些样品的缺陷短波单胞菌截留率。
结果示于下表II中。
表II
电纺纳米纤维垫的孔隙率越高,导致渗透率越高,同时仍然提供截留微生物的可靠手段。
使用方法
本发明所述的包含电纺纳米纤维的液体过滤介质用于食品业、饮料业、制药业、生物技术业、微电子业、化学处理、水处理及其它液体处理业中。
本发明所述的包含电纺纳米纤维的液体过滤介质可用于从液体样品或液体流过滤,分离,鉴定和/或检测微生物。
本发明所述的包含电纺纳米纤维的液体过滤介质可与任何液体样品制备方法一起使用,所述液体样品制备方法包括但不限于:色谱法;高压液相色谱法(HPLC);电泳;凝胶过滤;样品离心;在线样品制备;诊断试剂盒测试;诊断测试;高通量筛选;结合亲和力测定;液体样品纯化;依据尺寸分离流体样品的组分;依据物理性质分离流体样品的组分;依据化学性质分离流体样品的组分;依据生物性质分离流体样品的组分;依据静电性质分离流体样品的组分;以及它们的组合。另外,本发明所述的包含电纺纳米纤维的液体过滤介质可以是更大的装置和/或***的组件或部件。
配套装置
本发明还提供配套装置,其可用来从液体样品除去微生物。所述配套装置可包含,例如,一种或多种本发明所述的包含电纺纳米纤维的液体过滤介质,以及一个或多个液体过滤装置、所述介质的支持体或基材。所述配套装置可包含一种或多种对照物,并且可任选地包含用于实施本发明的方法的各种缓冲液。所述配套装置还可任选地包含用于清除试剂或非特异性截留的或结合的材料的清洗缓冲液。
其它任选的配套试剂包括洗脱缓冲液。所述各缓冲液可作为溶液在分开的容器中供应。或者,所述缓冲液可以以干燥的形式或粉末的形式提供,并且可根据用户期望的应用配制成溶液。在此情况中,所述缓冲液可以小包装中提供。在所述配套装置是自动化的并且是提供外力的手段如真空泵的情况中,所述装置可配备电源。所述配套装置还可包含说明书,用于指导使用包含液体过滤介质、装置、支持体或基材的电纺纳米纤维,和/或用于配制适合用于本发明的试剂,和实施本发明的方法。还可包含任选的软件,其用于在实施本发明的方法时或者在使用本发明的装置时记录和分析所得的数据。
术语"配套装置"包括,例如,合并在单个包装中的各组件,单独包装并一起销售的组件,或者一起出现在目录中的组件(例如,在目录中的相同页或跨页)。
上述公开可包括具有独立用途的多项不同的发明。虽然这些发明中的每项均以其优选形式被公开,但本文公开和所述的其具体实施方案不应理解为限制之意,因为许多改变是可能的。本发明的主题包括本文公开的各种要素、特征、功能和/或性质的所有新的非显而易见的组合和子组合。以下权利要求特别指出被认为是新的非显而易见的某些组合和子组合。体现在特征、功能、要素和/或性质的其它组合和子组合的发明可在要求本申请优先权的申请或相关申请中要求保护。所述权利要求,无论涉及不同的发明或者涉及相同的发明,无论就原权利要求而言更宽、更窄、相等或不同,也被认为涵盖在本公开的发明主题内。
Claims (10)
1.从液体样品中除去细菌的方法,其包括以下步骤:
a)提供包含微生物的液体样品,
b)提供多孔的包含纳米纤维的过滤介质,所述过滤介质的微生物对数下降值(LRV)大于约8,孔隙率为约80%-约95%,采用异丙醇测定的平均流量起泡点为1-100psi,并且液体渗透率大于约300LMH/psi,其中所述纳米纤维的纤维直径为约10nm-约1,000nm,和
c)使所述包含微生物的液体样品通过所述过滤介质,由此通过基于尺寸的分离来完全截留微生物。
2.权利要求1的方法,其中所述过滤介质的厚度为约1μm-约500μm。
3.权利要求1的方法,其中所述包含纳米纤维的过滤介质是通过选自电纺和电吹的方法制得的。
4.权利要求1的方法,其中所述细菌是支原体。
5.权利要求1的方法,其中所述纳米纤维包含聚合物,所述聚合物选自聚酰亚胺、脂肪族聚酰胺、芳香族聚酰胺、聚砜、乙酸纤维素酯、聚醚砜、聚氨酯、聚脲氨酯、聚苯并咪唑、聚醚酰亚胺、聚丙烯腈、聚对苯二甲酸乙二醇酯、聚丙烯、聚苯胺、聚氧化乙烯、聚萘二甲酸乙二醇酯、聚对苯二甲酸丁二醇酯、苯乙烯-丁二烯橡胶、聚苯乙烯、聚氯乙烯、聚乙烯醇、聚偏二氟乙烯、聚乙烯基丁烯、及它们的共聚物、衍生化合物或混合物。
6.权利要求1的方法,其中所述纳米纤维包含脂肪族聚酰胺。
7.权利要求1的方法,其中所述纳米纤维包含聚合物的混合物或共聚物的混合物。
8.权利要求1的方法,其中所述纳米纤维位于多孔支持体上。
9.权利要求8的方法,其中所述多孔支持体包含选自以下物质的一层或多层:纺粘无纺物、熔喷无纺物、针刺无纺物、水刺无纺物、湿法无纺物、树脂粘合无纺物、织造织物、针织织物、纸、及它们的组合。
10.权利要求4的方法,其中在用拉氏无胆甾原体进行挑战试验后,当所述过滤介质可被验证产生无菌流出液时,所述过滤介质完全截留支原体。
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CN102405061A (zh) | 2012-04-04 |
US9943616B2 (en) | 2018-04-17 |
JP2021007945A (ja) | 2021-01-28 |
JP6832889B2 (ja) | 2021-02-24 |
JP2018167264A (ja) | 2018-11-01 |
EP3381476A1 (en) | 2018-10-03 |
US20190015533A1 (en) | 2019-01-17 |
JP2014208342A (ja) | 2014-11-06 |
US9750829B2 (en) | 2017-09-05 |
JP2016185540A (ja) | 2016-10-27 |
WO2010107503A1 (en) | 2010-09-23 |
US20170360969A1 (en) | 2017-12-21 |
CN102405061B (zh) | 2015-12-09 |
JP2012520761A (ja) | 2012-09-10 |
US10722602B2 (en) | 2020-07-28 |
JP5931118B2 (ja) | 2016-06-08 |
US20170360970A1 (en) | 2017-12-21 |
US20120091072A1 (en) | 2012-04-19 |
US9889214B2 (en) | 2018-02-13 |
US20170360971A1 (en) | 2017-12-21 |
SG174346A1 (en) | 2011-11-28 |
US10064965B2 (en) | 2018-09-04 |
EP2408482A1 (en) | 2012-01-25 |
ES2932986T3 (es) | 2023-01-30 |
JP7100099B2 (ja) | 2022-07-12 |
SG10201801667YA (en) | 2018-03-28 |
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EP3381476B1 (en) | 2022-11-09 |
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