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    Quantitative cell analysis methods employing magnetic separation
    4.
    发明授权
    Quantitative cell analysis methods employing magnetic separation 失效
    使用磁分离的定量细胞分析方法

    公开(公告)号：US5993665A

    公开(公告)日：1999-11-30

    申请号：US867008

    申请日：1997-06-02

    申请人： Leon W. M. M. Terstappen ,  Paul A. Liberti

    发明人： Leon W. M. M. Terstappen ,  Paul A. Liberti

    IPC分类号： B03C1/025 ,  B01D35/06 ,  B03C1/034 ,  B03C1/035 ,  G01N33/533 ,  G01N33/543 ,  B01D35/00

    CPC分类号： B03C1/025 ,  B01L3/502 ,  B03C1/034 ,  B03C1/035 ,  G01N27/745 ,  G01N33/54326 ,  G01N33/54366 ,  B01L2400/043 ,  Y10T436/108331 ,  Y10T436/25375

    摘要： A method of quantitative analysis of microscopic biological specimens in a fluid medium is disclosed in which the specimens are rendered magnetically responsive by immunospecific binding with ferromagnetic colloid. A known quantity of magnetically-responsive marker particles are added to the fluid medium. The fluid medium is then subjected to a magnetic separation process, to collect the magnetic species from the fluid. The collected species are resuspended in a second fluid medium, and the relative quantities thereof are enumerated to determine the concentration of the desired biological specimen in the first fluid medium. The marker particles may comprise magnetic particles having a relatively large magnetic moment, a magnetic moment approximately equal to the magnetically-labelled biological speciment of interest, or both in order to compensate the determination for variations in immunospecific binding affinity and/or magnetic collection efficiency. The present method is useful for conducting hematological assays, such as red blood cell counts and related determinations. In the case of red blood cells, the specimens may be rendered magnetically responsive by reduction of the iron atoms present in hemoglobin. The enumeration step of the method may be conducted with such equipment as a flow cytometer, or by a novel optical scanning magnetic cell immobilization apparatus.

    摘要翻译： 公开了一种流体介质中微观生物样本的定量分析方法，其中通过与铁磁性胶体的免疫特异性结合使样本呈现磁响应。 将已知量的磁响应标记颗粒添加到流体介质中。 然后对流体介质进行磁分离处理，以从流体中收集磁性物质。 将收集的物质重新悬浮在第二流体培养基中，并列举其相对量以确定第一流体培养基中所需生物样品的浓度。 标记颗粒可以包括具有相对大的磁矩，大约等于感兴趣的磁性标记的生物学物理学的磁矩或两者的磁性颗粒，以补偿免疫特异性结合亲和力和/或磁收集效率的变化的确定。 本方法可用于进行血液学测定，如红细胞计数和相关测定。 在红细胞的情况下，可以通过还原血红蛋白中存在的铁原子来使样品呈现磁响应性。 该方法的计数步骤可以用流式细胞仪等设备进行，也可以通过新的光学扫描磁性细胞固定装置进行。

    Affinity-binding separation and release of one or more selected subset of biological entities from a mixed population thereof
    5.
    发明授权
    Affinity-binding separation and release of one or more selected subset of biological entities from a mixed population thereof 失效
    从其混合群体中亲和力结合分离和释放一个或多个选定的生物实体子集

    公开(公告)号：US5646001A

    公开(公告)日：1997-07-08

    申请号：US395967

    申请日：1995-02-28

    申请人： Leon W. M. M. Terstappen ,  Galla C. Rao ,  Dhanesh I. Gohel ,  Brian P. Feeley ,  Steven Gross ,  Ellen S. Church ,  Paul A. Liberti

    发明人： Leon W. M. M. Terstappen ,  Galla C. Rao ,  Dhanesh I. Gohel ,  Brian P. Feeley ,  Steven Gross ,  Ellen S. Church ,  Paul A. Liberti

    IPC分类号： A23L3/32 ,  B03C1/035 ,  G01N33/543 ,  G01N33/569 ,  G01N35/00 ,  G01N33/567

    CPC分类号： A23L3/32 ,  B03C1/035 ,  G01N33/54326 ,  G01N33/569 ,  G01N35/0098 ,  Y10S435/962 ,  Y10S435/973 ,  Y10S436/824 ,  Y10S436/825 ,  Y10T436/25

    摘要： A method for separation of a mixture of biological entities into at least three distinct, subpopulations. Different antibodies are provided, with each antibody bound to a solid support in a unique manner such that by a manipulation of the physical or chemical environment, the bonds between the antibodies and the solid supports can be selectively broken. The mixed population of cells is incubated with the antibodies. The cells are magnetically separated from a test medium and collected in a monolayer upon a collection surface. Then by manipulation of the physicochemical environment, specific linkages can be broken and desired cell subpopulations released from the collection surface. This method has medically significant diagnostic and therapeutic applications, as entire cell types can be separated from non-malignant medically vital cell types. Cancer can be diagnosed, staged, and monitored. Genetic analysis from maternal blood, CVS, or amniocentesis samples is possible. Diseases such as AIDS, tuberculosis or hepatitis can be monitored. This invention also has utility in the fields of bone marrow transplantation, fetal cell research, in vitro fertilization, and gene therapy.

    摘要翻译： 将生物实体的混合物分离成至少三个不同的亚群体的方法。 提供了不同的抗体，每种抗体以独特的方式结合到固体支持物上，使得通过操纵物理或化学环境，可以选择性地破坏抗体和固体支持物之间的键。 将混合的细胞群与抗体一起孵育。 将细胞与测试介质磁分离并收集在收集表面上的单层中。 然后通过操纵物理化学环境，可以破坏特定的连接并且从收集表面释放所需的细胞亚群。 该方法具有医学上显着的诊断和治疗应用，因为整个细胞类型可以与非恶性的医学上重要的细胞类型分离。 可以诊断，分期和监测癌症。 从母体血液，CVS或羊膜穿刺样品的遗传分析是可能的。 可以监测艾滋病，结核病或肝炎等疾病。 本发明还可用于骨髓移植，胎儿细胞研究，体外受精和基因治疗领域。

    Methods for biological substance analysis employing internal magnetic gradients separation and an externally-applied transport force
    8.
    发明授权
    Methods for biological substance analysis employing internal magnetic gradients separation and an externally-applied transport force 失效
    使用内部磁梯分离和外部施加的输送力进行生物物质分析的方法

    公开(公告)号：US6013188A

    公开(公告)日：2000-01-11

    申请号：US867233

    申请日：1997-06-02

    申请人： Leon W. M. M. Terstappen ,  Paul A. Liberti

    发明人： Leon W. M. M. Terstappen ,  Paul A. Liberti

    IPC分类号： B03C1/025 ,  B01D35/06 ,  B03C1/034 ,  B03C1/035 ,  G01N33/533 ,  G01N33/543

    CPC分类号： B03C1/025 ,  B01L3/502 ,  B03C1/034 ,  B03C1/035 ,  G01N27/745 ,  G01N33/54326 ,  G01N33/54366 ,  B01L2400/043 ,  Y10T436/108331 ,  Y10T436/25375

    摘要： A method of qualitative analysis of microscopic biological specimens is disclosed in which the specimens are rendered magnetically responsive by immunosspecific binding with feuromagnetic colloid. The specimens are placed in a vessel having a feuromagnetic capture structure supported on a transparent wall. The vessel is then placed in a magnetic field for inducing a high gradient region in the vicinity of the capture structure and, in particular, adjacent to the capture structure along the direction of the applied field. The magnetically responsive specimens are collected and immobilized by the high gradient along the wall adjacent to the capture structure for unobstructed microscopic observation. Subpapulations of distinct types of cells can thus be observed and differentiated. As transport force, such as gravity or an externally applied magnetic gradient, may be applied to the vessel to move the specimens into the high gradient region.

    摘要翻译： 公开了一种微观生物样品的定性分析方法，其中通过与feuromagnetic胶体的免疫特异性结合使样品呈现磁响应。 将样品放置在具有支撑在透明壁上的feuromagnetic捕获结构的容器中。 然后将容器放置在磁场中，以在捕获结构附近诱导高梯度区域，并且特别是沿着施加场的方向与捕获结构相邻。 通过与捕获结构相邻的壁的高梯度收集和固定磁响应样品，用于无阻碍的显微镜观察。 因此可以观察和区分不同类型细胞的突变。 由于诸如重力或外部施加的磁梯度的输送力可以施加到容器以将样品移动到高梯度区域。

    Surface immobilization of magnetically collected materials
    9.
    发明授权
    Surface immobilization of magnetically collected materials 失效
    磁性收集材料的表面固定

    公开(公告)号：US5660990A

    公开(公告)日：1997-08-26

    申请号：US516694

    申请日：1995-08-18

    申请人： Galla Chandra Rao ,  Paul A. Liberti

    发明人： Galla Chandra Rao ,  Paul A. Liberti

    IPC分类号： C12Q1/68 ,  G01N33/543 ,  C12Q1/70 ,  G01N33/53

    CPC分类号： C12Q1/6813 ,  C12Q1/6834 ,  G01N33/54313 ,  Y10S436/807 ,  Y10S436/824 ,  Y10S977/838 ,  Y10S977/853 ,  Y10S977/918 ,  Y10S977/958 ,  Y10S977/96

    摘要： A method for determining the presence and/or concentration of a target substance e.g. protein, nucleic acid, bioparticle etc. in a fluid sample is provided. The method disclosed combines elements of immunoassays, coated cup assays and magnetic particle separation to effect the quantitation and recovery of an analyte in solution. Also the method ensures the non-reorientation of magnetically collected material by linking the magnetic particles to a collection surface via a specific binding pair. This linkage immobilizes the magnetic-analyte-containing material and thus allows for vigorous washing and reagent addition without significant redistribution or displacement. Thus the assay of this invention offers the speed of diffusion controlled kinetics as in a ferrofluid assay, the speed of collection of labeled target substance as in a magnetic assay as well as the ability to magnetically monolayer the ferrofluid, all of which is combined with the ease of washing and signal detection found in a coated cup assay.

    摘要翻译： 用于确定靶物质的存在和/或浓度的方法，例如， 提供了流体样品中的蛋白质，核酸，生物颗粒等。 所公开的方法组合了免疫测定，涂覆杯测定和磁性颗粒分离的元件，以实现溶液中分析物的定量和回收。 此外，该方法通过经由特异性结合对将磁性颗粒连接到收集表面来确保磁性收集材料的非重新取向。 该连接固定了含有磁性分析物的材料，从而允许剧烈的洗涤和试剂添加，而不会有显着的再分配或位移。 因此，本发明的测定提供了如铁磁流体测定中的扩散控制动力学速度，如在磁性测定中收集标记的靶物质的速度以及磁性单层铁磁流体的能力，所有这些都与 在涂层杯测定中发现易洗涤和信号检测。

    Magnetic immobilization and manipulation of biological entities
    10.
    发明授权
    Magnetic immobilization and manipulation of biological entities 失效
    磁固定和操纵生物实体

    公开(公告)号：US5876593A

    公开(公告)日：1999-03-02

    申请号：US931067

    申请日：1997-09-15

    申请人： Paul A. Liberti ,  Yuzhou Wang

    发明人： Paul A. Liberti ,  Yuzhou Wang

    IPC分类号： B01D35/06 ,  B03C1/00 ,  B03C1/01 ,  B03C1/025 ,  B03C1/032 ,  B03C1/033 ,  B03C1/034 ,  B03C1/035 ,  B03C1/28 ,  C12M3/00 ,  G01N33/483 ,  G01N33/543 ,  G01N33/569 ,  G01N35/00 ,  G01N35/02

    CPC分类号： B03C1/002 ,  B03C1/01 ,  B03C1/025 ,  B03C1/032 ,  B03C1/0332 ,  B03C1/034 ,  B03C1/035 ,  C12M47/04 ,  G01N33/54326 ,  G01N33/56972 ,  G01N35/026 ,  G01N2333/70589 ,  G01N35/0098 ,  Y10T436/108331 ,  Y10T436/25375

    摘要： Biological entities such as cells, microbes, or components thereof are labeled with a magnetic colloid containing microscopic magnetic particles. The magnetic particles have a coating capable of biospecific or non-specific binding with the entities. An immobilization apparatus includes a non-magnetic vessel having a ferromagnetic collection structure for attracting the entities toward a collection surface upon which the magnetically labeled entities are immobilized subsequent to placement of the vessel on a support between two magnets. The ferromagnetic collection structure preferably has a sharp edge or high curvature for intensifying the magnetic field and for collecting the entities in a monolayer. The vessel includes an un-obstructed observation path so that immobilized entities may be observed and/or manipulated. The ferromagnetic collection structure may be arranged in various two dimensional patterns to provide a desired collection configuration. The apparatus may further have inlet and outlet ports for allowing a flow of liquid reagent through the vessel for washing or straining the immobilized entities. The support can be translated so that a shoulder on the collection structure concentrates the collected entities by movement of the shoulder in a transverse direction relative to the magnetic field.

    摘要翻译： PCT No.PCT / US93 / 11087 Sec。 371日期1995年04月24日 102（e）1995年4月24日PCT PCT 1997年9月15日PCT公布。 公开号WO94 / 11078 日期1994年5月26日生物实体如细胞，微生物或其组分用含有微观磁性颗粒的磁性胶体标记。 磁性颗粒具有能够与实体进行生物特异性或非特异性结合的涂层。 固定装置包括具有铁磁收集结构的非磁性容器，用于在将容器放置在两个磁体之间的支撑件上时，将实体吸引到收集表面，在该收集表面上磁性标记的实体被固定。 铁磁收集结构优选地具有尖锐的边缘或高曲率，用于增强磁场和用于在单层中收集实体。 容器包括未阻挡的观察路径，使得可以观察和/或操纵固定的实体。 铁磁收集结构可以以各种二维图案布置以提供期望的收集配置。 该装置还可以具有用于允许液体试剂流过容器的入口和出口，用于洗涤或使固定的实体变窄。 支撑件可以平移，使得收集结构上的肩部通过肩部相对于磁场的横向方向的移动来集中收集的实体。