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    Protein fragment complementation assays for high-throughput and high-content screening
    1.
    发明授权
    Protein fragment complementation assays for high-throughput and high-content screening 失效
    用于高通量和高含量筛选的蛋白质片段互补测定

    公开(公告)号：US07935493B2

    公开(公告)日：2011-05-03

    申请号：US11450379

    申请日：2006-06-12

    申请人： Stephen William Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John K. Westwick

    发明人： Stephen William Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John K. Westwick

    IPC分类号： G01N33/53 ,  G01N33/532 ,  G01N33/533 ,  G01N33/573 ,  G01N21/00 ,  G01N21/76 ,  C07K14/00

    CPC分类号： G01N33/5011 ,  G01N33/5008 ,  G01N33/5041 ,  G01N33/542 ,  G01N33/6845 ,  G01N2500/02 ,  Y02A90/26

    摘要： The present invention provides protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. The interacting pair may be selected by cDNA library screening; by gene-by-gene interaction mapping; or by prior knowledge of a pathway. Fluorescent and luminescent assays can be constructed using the methods provided herein. The selection of suitable PCA reporters for high-throughput or high-content (high-context) assay formats is described for a diversity of reporters, with particular detail provided for examples of monomeric enzymes and fluorescent proteins. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. Single-color and multi-color assays are disclosed. Further disclosed are universal expression vectors with cassettes that allow the rapid construction of assays for a large and diverse number of gene/reporter combinations. The development of such assays is shown to be straightforward, providing for a broad, flexible and biologically relevant platform for drug discovery.

    摘要翻译： 本发明提供用于药物发现的蛋白质片段互补测定法，特别是鉴定激活或抑制细胞途径的化合物。 基于与适当的PCA报道子组合的相互作用的蛋白质对的选择，测定可以以高通量或高含量模式进行，并且可以用于化合物文库的自动化筛选。 可以通过cDNA文库筛选来选择相互作用的对; 通过基因相互作用作图; 或通过路径的先验知识。 可以使用本文提供的方法构建荧光和发光测定。 针对多种报道者描述了适合于高通量或高含量（高上下文）测定形式的PCA报告人的选择，具体提供了单体酶和荧光蛋白的实例。 描述了用于构建生物化学途径中的一个或多个步骤的这种测定方法; 测试来自组合，天然产物，肽，抗体，核酸或其他不同文库的化合物对感兴趣的蛋白质或途径的影响; 并使用筛选结果来鉴定激活或抑制感兴趣的蛋白质或途径的特定化合物。 公开了单色和多色测定。 进一步公开的是具有盒的通用表达载体，其允许快速构建用于大量和多样化数量的基因/报道子组合的测定。 显示出这种测定的发展是直接的，为药物发现提供了广泛，灵活和生物学上相关的平台。

    Protein fragment complementation assays for high-throughput and high-content screening
    2.
    发明申请
    Protein fragment complementation assays for high-throughput and high-content screening 审中-公开
    用于高通量和高含量筛选的蛋白质片段互补测定

    公开(公告)号：US20120149597A1

    公开(公告)日：2012-06-14

    申请号：US13067007

    申请日：2011-05-02

    申请人： Stephen William Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John K. Westwick

    发明人： Stephen William Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John K. Westwick

    IPC分类号： C40B30/06 ,  C40B40/10

    CPC分类号： G01N33/5011 ,  G01N33/5008 ,  G01N33/5041 ,  G01N33/542 ,  G01N33/6845 ,  G01N2500/02 ,  Y02A90/26

    摘要： The present invention provides protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. The interacting pair may be selected by cDNA library screening; by gene-by-gene interaction mapping; or by prior knowledge of a pathway. Fluorescent and luminescent assays can be constructed using the methods provided herein. The selection of suitable PCA reporters for high-throughput or high-content (high-context) assay formats is described for a diversity of reporters, with particular detail provided for examples of monomeric enzymes and fluorescent proteins. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. Single-color and multi-color assays are disclosed. Further disclosed are universal expression vectors with cassettes that allow the rapid construction of assays for a large and diverse number of gene/reporter combinations. The development of such assays is shown to be straightforward, providing for a broad, flexible and biologically relevant platform for drug discovery.

    摘要翻译： 本发明提供用于药物发现的蛋白质片段互补测定法，特别是鉴定激活或抑制细胞途径的化合物。 基于与适当的PCA报道子组合的相互作用的蛋白质对的选择，测定可以以高通量或高含量模式进行，并且可以用于化合物文库的自动化筛选。 可以通过cDNA文库筛选来选择相互作用的对; 通过基因相互作用作图; 或通过路径的先验知识。 可以使用本文提供的方法构建荧光和发光测定。 针对多种报道者描述了适合于高通量或高含量（高上下文）测定形式的PCA报告人的选择，具体提供了单体酶和荧光蛋白的实例。 描述了用于构建生物化学途径中的一个或多个步骤的这种测定方法; 测试来自组合，天然产物，肽，抗体，核酸或其他不同文库的化合物对感兴趣的蛋白质或途径的影响; 并使用筛选结果来鉴定激活或抑制感兴趣的蛋白质或途径的特定化合物。 公开了单色和多色测定。 进一步公开的是具有盒的通用表达载体，其允许快速构建用于大量和多样化数量的基因/报道子组合的测定。 显示出这种测定的发展是直接的，为药物发现提供了广泛，灵活和生物相关的平台。

    Protein fragment complementation assays for high-throughput and high-content screening
    3.
    发明授权
    Protein fragment complementation assays for high-throughput and high-content screening 失效
    用于高通量和高含量筛选的蛋白质片段互补测定

    公开(公告)号：US07062219B2

    公开(公告)日：2006-06-13

    申请号：US10772021

    申请日：2004-02-05

    申请人： Stephen William Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John K. Westwick

    发明人： Stephen William Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John K. Westwick

    IPC分类号： C12Q1/00 ,  C07K14/00 ,  C12N15/11

    CPC分类号： G01N33/5011 ,  G01N33/5008 ,  G01N33/5041 ,  G01N33/542 ,  G01N33/6845 ,  G01N2500/02 ,  Y02A90/26

    摘要： The present invention provides protein single-color and multi-color protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter such as monomeric enzymes and fluorescent proteins, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. The development of such assays provides for a broad, flexible and biologically relevant platform for drug discovery.

    摘要翻译： 本发明提供用于药物发现的蛋白质单色和多色蛋白质片段互补测定法，特别是鉴定激活或抑制细胞途径的化合物。 基于与适当的PCA报告物如单体酶和荧光蛋白结合的相互作用的蛋白质对的选择，测定可以以高通量或高含量模式进行，并且可以用于化合物文库的自动筛选。 描述了用于构建生物化学途径中的一个或多个步骤的这种测定方法; 测试来自组合，天然产物，肽，抗体，核酸或其他不同文库的化合物对感兴趣的蛋白质或途径的影响; 并使用筛选结果来鉴定激活或抑制感兴趣的蛋白质或途径的特定化合物。 这种测定的开发提供了用于药物发现的广泛，灵活和生物相关的平台。

    Protein fragment complementation assays for high-throughput and high-content screening
    4.
    发明申请
    Protein fragment complementation assays for high-throughput and high-content screening 失效
    用于高通量和高含量筛选的蛋白质片段互补测定

    公开(公告)号：US20060224331A1

    公开(公告)日：2006-10-05

    申请号：US11450379

    申请日：2006-06-12

    申请人： Stephen Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John Westwick

    发明人： Stephen Watson Michnick ,  Ingrid Remy ,  Marnie MacDonald ,  Jane Lamerdin ,  Helen Yu ,  John Westwick

    IPC分类号： C40B30/02 ,  G06F19/00

    CPC分类号： G01N33/5011 ,  G01N33/5008 ,  G01N33/5041 ,  G01N33/542 ,  G01N33/6845 ,  G01N2500/02 ,  Y02A90/26

    摘要： The present invention provides protein fragment complementation assays for drug discovery, in particular to identify compounds that activate or inhibit cellular pathways. Based on the selection of an interacting protein pair combined with an appropriate PCA reporter, the assays may be run in high-throughput or high-content mode and may be used in automated screening of libraries of compounds. The interacting pair may be selected by cDNA library screening; by gene-by-gene interaction mapping; or by prior knowledge of a pathway. Fluorescent and luminescent assays can be constructed using the methods provided herein. The selection of suitable PCA reporters for high-throughput or high-content (high-context) assay formats is described for a diversity of reporters, with particular detail provided for examples of monomeric enzymes and fluorescent proteins. Methods are described for constructing such assays for one or more steps in a biochemical pathway; testing the effects of compounds from combinatorial, natural product, peptide, antibody, nucleic acid or other diverse libraries on the protein or pathway(s) of interest; and using the results of the screening to identify specific compounds that activate or inhibit the protein or pathway(s) of interest. Single-color and multi-color assays are disclosed. Further disclosed are universal expression vectors with cassettes that allow the rapid construction of assays for a large and diverse number of gene/reporter combinations. The development of such assays is shown to be straightforward, providing for a broad, flexible and biologically relevant platform for drug discovery.

    摘要翻译： 本发明提供用于药物发现的蛋白质片段互补测定法，特别是鉴定激活或抑制细胞途径的化合物。 基于与适当的PCA报道子组合的相互作用的蛋白质对的选择，测定可以以高通量或高含量模式进行，并且可以用于化合物文库的自动化筛选。 可以通过cDNA文库筛选来选择相互作用的对; 通过基因相互作用作图; 或通过路径的先验知识。 可以使用本文提供的方法构建荧光和发光测定。 针对多种报道者描述了适合于高通量或高含量（高上下文）测定形式的PCA报告人的选择，具体提供了单体酶和荧光蛋白的实例。 描述了用于构建生物化学途径中的一个或多个步骤的这种测定方法; 测试来自组合，天然产物，肽，抗体，核酸或其他不同文库的化合物对感兴趣的蛋白质或途径的影响; 并使用筛选结果来鉴定激活或抑制感兴趣的蛋白质或途径的特定化合物。 公开了单色和多色测定。 进一步公开的是具有盒的通用表达载体，其允许快速构建用于大量和多样化数量的基因/报道子组合的测定。 显示出这种测定的发展是直接的，为药物发现提供了广泛，灵活和生物相关的平台。

    In vivo screening of protein-protein interactions with protein-fragment complementation assays
    5.
    发明授权
    In vivo screening of protein-protein interactions with protein-fragment complementation assays 失效
    蛋白质 - 蛋白质相互作用与蛋白质片段互补测定的体内筛选

    公开(公告)号：US07855167B2

    公开(公告)日：2010-12-21

    申请号：US10728355

    申请日：2003-12-05

    申请人： Stephen William Watson Michnick ,  Ingrid Remy ,  Jane Lamerdin

    发明人： Stephen William Watson Michnick ,  Ingrid Remy ,  Jane Lamerdin

    IPC分类号： C40B20/08 ,  C40B30/10 ,  C12Q1/70 ,  C12P21/04 ,  C07H21/02 ,  C07H21/04

    CPC分类号： G01N33/542 ,  C07K14/43595 ,  G01N33/58 ,  G01N33/68 ,  G01N2800/52

    摘要： The present invention describes rapid methods to screen for biomolecular interactions in vivo based on protein fragment complementation assays (PCA). We have demonstrated an in vivo library-versus-library screening strategy that has numerous applications in the identification of novel protein-protein interactions and in directed evolution. Also we demonstrate the detection of protein-protein interactions starting with defined (full-length) cDNAs, and the concomitant generation of functional assays that provide initial validation of the cDNA products as being biologically relevant. Also, we screened a large cDNA collection using automated PCA, combined with quantitative detection of protein-protein complexes. The invention enables bait-vs.-library, library-vs.-library and defined gene screening in any type of cell or cellular context, and using a wide range of reporters and detection methods. The invention allows for identifying and validating genes involved in any cellular process and also provide assays to study effects of potential drugs, or gene knockouts on specific pathways.

    摘要翻译： 本发明描述了基于蛋白质片段互补测定（PCA）在体内筛选生物分子相互作用的快速方法。 我们已经展示了体内文库对文库筛选策略，其在鉴定新的蛋白质 - 蛋白质相互作用和定向进化中具有许多应用。 我们还展示了从定义的（全长）cDNA开始的蛋白质 - 蛋白质相互作用的检测，以及伴随产生的功能测定，提供cDNA产物的初步验证是生物相关的。 此外，我们使用自动PCA筛选了大量cDNA集合，结合蛋白质 - 蛋白质复合物的定量检测。 本发明使得在任何类型的细胞或细胞上下文中使用诱饵与文库，文库与文库以及定义的基因筛选，以及使用广泛的记者和检测方法。 本发明允许鉴定和验证参与任何细胞过程的基因，并且还提供测定以研究潜在药物或基因敲除对特定途径的影响。

    Protein fragment complementation assays for the detection of biological or drug interactions
    6.
    发明申请
    Protein fragment complementation assays for the detection of biological or drug interactions 有权
    用于检测生物或药物相互作用的蛋白质片段互补测定法

    公开(公告)号：US20110287950A1

    公开(公告)日：2011-11-24

    申请号：US13137257

    申请日：2011-08-01

    申请人： Stephen William Watson Michnick ,  Joelle Nina Polletier ,  Ingrid Remy

    发明人： Stephen William Watson Michnick ,  Joelle Nina Polletier ,  Ingrid Remy

    IPC分类号： C40B30/00 ,  C07K14/34 ,  C07K14/435 ,  C12N9/96 ,  C12Q1/66 ,  C12Q1/48 ,  C12Q1/34 ,  C12Q1/26 ,  C07K14/21 ,  C07K14/00

    CPC分类号： G01N33/6845 ,  A01K67/0271 ,  A01K2267/0331 ,  A01K2267/0393 ,  C07K14/43595 ,  C07K2319/00 ,  C12N15/1055 ,  G01N33/5008 ,  G01N33/542 ,  G01N33/573 ,  G01N33/58 ,  G01N33/581 ,  G01N33/68 ,  G01N33/6803 ,  Y02A90/26

    摘要： We describe a strategy for designing and implementing protein-fragment complementation assays (PCAs) to detect biomolecular interactions in vivo and in vitro. The design, implementation and broad applications of this strategy are illustrated with a large number of enzymes with particular detail provided for the example of murine dihydrofolate reductase (DHFR). Fusion peptides consisting of N- and C-terminal fragments of murine DHFR fused to GCN4 leucine zipper sequences were coexpressed in Escherichia coli grown in minimal medium, where the endogenous DHFR activity was inhibited with trimethoprim. Coexpression of the complementary fusion products restored colony formation. Survival only occurred when both DHFR fragments were present and contained leucine-zipper forming sequences, demonstrating that reconstitution of enzyme activity requires assistance of leucine zipper formation. DHFR fragment-interface point mutants of increasing severity (Ile to Val, Ala and Gly) resulted in a sequential increase in E. coli doubling times illustrating the successful DHFR fragment reassembly rather than non-specific interactions between fragments. This assay could be used to study equilibrium and kinetic aspects of molecular interactions including protein-protein, protein-DNA, protein-RNA, protein-carbohydrate and protein-small molecule interactions, for screening cDNA libraries for binding of a target protein with unknown proteins or libraries of small organic molecules for biological activity. The selection and design criteria applied here is developed for numerous examples of clonal selection, colorometric, fluorometric and other assays based on enzymes whose products can be measured. The development of such assay systems is shown to be simple, and provides for a diverse set of protein fragment complementation applications.

    摘要翻译： 我们描述了设计和实施蛋白质片段互补测定（PCAs）以检测体内和体外生物分子相互作用的策略。 该策略的设计，实现和广泛应用用大量酶进行了说明，具体细节提供了鼠二氢叶酸还原酶（DHFR）的例子。 由融合到GCN4亮氨酸拉链序列的鼠DHFR的N-和C-末端片段组成的融合肽在基本培养基中生长的大肠杆菌中共表达，其中内源性DHFR活性被甲氧苄啶抑制。 互补融合产物的共表达恢复了菌落形成。 存活只发生在两个DHFR片段存在并含有亮氨酸 - 拉链形成序列时，表明重组酶活性需要辅助亮氨酸拉链形成。 增加严重程度（Ile至Val，Ala和Gly）的DHFR片段 - 接口点突变体导致大肠杆菌倍增时间的顺序增加，说明成功的DHFR片段重组而不是片段之间的非特异性相互作用。 该测定可用于研究包括蛋白质蛋白质，蛋白质-DNA，蛋白质-RNA，蛋白质 - 碳水化合物和蛋白质 - 小分子相互作用在内的分子相互作用的平衡和动力学方面，用于筛选靶向蛋白质与未知蛋白质结合的cDNA文库 或用于生物活性的小有机分子的文库。 这里应用的选择和设计标准是针对克隆选择，色度，荧光测定和其他基于可以测量其产物的酶的测定的许多实例开发的。 这种测定系统的开发被证明是简单的，并且提供了多种蛋白质片段互补应用的集合。

    Protein fragment complementation assays for the detection of biological or drug interactions
    9.
    发明授权
    Protein fragment complementation assays for the detection of biological or drug interactions 失效
    用于检测生物或药物相互作用的蛋白质片段互补测定法

    公开(公告)号：US06428951B1

    公开(公告)日：2002-08-06

    申请号：US09499464

    申请日：2000-02-07

    申请人： Stephen William Watson Michnick ,  Joelle Nina Pelletier ,  Ingrid Remy

    发明人： Stephen William Watson Michnick ,  Joelle Nina Pelletier ,  Ingrid Remy

    IPC分类号： C12Q125

    CPC分类号： G01N33/6845 ,  A01K67/0271 ,  A01K2267/0331 ,  A01K2267/0393 ,  C07K14/43595 ,  C07K2319/00 ,  C12N15/1055 ,  G01N33/5008 ,  G01N33/542 ,  G01N33/573 ,  G01N33/58 ,  G01N33/581 ,  G01N33/68 ,  G01N33/6803 ,  Y02A90/26

    摘要： We describe a strategy for designing and implementing protein-fragment complementation assays (PCAs) to detect biomolecular interactions in vivo and in vitro. The design, implementation and broad applications of this strategy are illustrated with a large number of enzymes with particular detail provided for the example of murine dihydrofolate reductase (DHFR). Fusion peptides consisting of N- and C-terminal fragments of murine DHFR fused to-GCN4 leucine zipper sequences were coexpressed in Escherichia coli grown in minimal medium, where the endogenous DHFR activity was inhibited with trimethoprim. Coexpression of the complementary fusion products restored colony formation. Survival only occurred when both DHFR fragments were present and contained leucine-zipper forming sequences, demonstrating that reconstitution of enzyme activity requires assistance of leucine zipper formation. DHFR fragment-interface point mutants of increasing severity (lie to Val, Ala and Gly) resulted in a sequential increase in E. coli doubling times illustrating the successful DHFR fragment reassembly rather than non-specific interactions between fragments. This assay could be used to study equilibrium and kinetic aspects of molecular interactions including protein-protein, protein-DNA, protein-RNA, protein-carbohydrate and protein-small molecule interactions, for screening cDNA libraries for binding of a target protein with unknown proteins or libraries of small organic molecules for biological activity. The selection and design criteria applied here is developed for numerous examples of clonal selection, colorometric, fluorometric and other assays based on enzymes whose products can be measured. The development of such assay systems is shown to be simple, and provides for a diverse set of protein fragment complementation applications.

    摘要翻译： 我们描述了设计和实施蛋白质片段互补测定（PCAs）以检测体内和体外生物分子相互作用的策略。 该策略的设计，实现和广泛应用用大量酶进行了说明，具体细节提供了鼠二氢叶酸还原酶（DHFR）的例子。 由小鼠DHFR融合到GCN4亮氨酸拉链序列的N-和C-末端片段组成的融合肽在基本培养基中生长的大肠杆菌中共表达，其中内源性DHFR活性被甲氧苄啶抑制。 互补融合产物的共表达恢复了菌落形成。 存活只发生在两个DHFR片段存在并含有亮氨酸 - 拉链形成序列时，表明重组酶活性需要辅助亮氨酸拉链形成。 增加严重程度的DHFR片段 - 接口点突变体（以Val，Ala和Gly表示）导致大肠杆菌倍增时间顺序增加，说明成功的DHFR片段重组而不是片段之间的非特异性相互作用。 该测定可用于研究包括蛋白质蛋白质，蛋白质-DNA，蛋白质-RNA，蛋白质 - 碳水化合物和蛋白质 - 小分子相互作用在内的分子相互作用的平衡和动力学方面，用于筛选靶向蛋白质与未知蛋白质结合的cDNA文库 或用于生物活性的小有机分子的文库。 这里应用的选择和设计标准是针对克隆选择，色度，荧光测定和其他基于可以测量其产物的酶的测定的许多实例开发的。 这种测定系统的开发被证明是简单的，并且提供了多种蛋白质片段互补应用的集合。

    Protein fragment complementation assays for the detection of biological or drug interactions
    10.
    发明授权
    Protein fragment complementation assays for the detection of biological or drug interactions 失效
    用于检测生物或药物相互作用的蛋白质片段互补测定法

    公开(公告)号：US06294330B1

    公开(公告)日：2001-09-25

    申请号：US09124850

    申请日：1998-07-30

    申请人： Stephen William Watson Michnick ,  Ingrid Remy

    发明人： Stephen William Watson Michnick ,  Ingrid Remy

    IPC分类号： C12Q168

    CPC分类号： C07K14/43595 ,  C07K2319/00 ,  C12N15/1055 ,  G01N33/542 ,  G01N33/573 ,  G01N33/58 ,  G01N33/581 ,  G01N33/68 ,  G01N33/6803 ,  G01N33/6845 ,  Y02A90/26

    摘要： The invention provides a general protein-fragment complementation assays to detect biomolecular interactions in vivo and in vitro. The protein-complemetation assay/universal reporter system can be used to detect and screen an agonist and an antagonist of a membrane receptor system. The assay can be used to study protein-protein, protein-DNA, protein-RNA, protein-carbohydrate, and protein-small molecule interactions. The assay can be used to screen cDNA libraries for binding of a target protein with unknown proteins or libraries of small organic molecules for biological activity.

    摘要翻译： 本发明提供了在体内和体外检测生物分子相互作用的一般蛋白质片段互补测定法。 蛋白质完全测定/通用报道系统可用于检测和筛选膜受体系统的激动剂和拮抗剂。 该测定法可用于研究蛋白质 - 蛋白质，蛋白质 - DNA，蛋白质 - RNA，蛋白质 - 碳水化合物和蛋白质 - 小分子相互作用。 该测定法可用于筛选cDNA文库以将靶蛋白与未知蛋白质或小有机分子的文库结合用于生物活性。