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    • 4. 发明申请
    • Efficient generation of expression cell lines through the use of scorable homeostatic reporter genes
    • 通过使用可扫描的稳态报告基因,有效产生表达细胞系
    • US20060286671A1
    • 2006-12-21
    • US11509177
    • 2006-08-23
    • Robert DuBridge
    • Robert DuBridge
    • C12N15/74C12N15/09
    • C07K16/00C12N15/907C12N2800/30C12N2840/203C12Q1/6897
    • The present invention provides methods for site-specific recombination in a cell, as well as vectors which can be employed in such methods. The methods and vectors of the present invention can be used to obtain persistent gene expression in a cell and to modulate gene expression. One preferred method according to the invention comprises contacting a cell with a vector comprising an origin of replication functional in mammalian cells located between first and second recombining sites located in parallel. Another preferred method comprises, in part, contacting a cell with a vector comprising first and second recombining sites in antiparallel orientations such that the vector is internalized by the cell. In both methods, the cell is further provided with a site-specific recombinase that effects recombination between the first and second recombining sites of the vector.
    • 本发明提供了细胞中位点特异性重组的方法,以及可用于这些方法的载体。 本发明的方法和载体可用于获得细胞中持续的基因表达并调节基因表达。 根据本发明的一种优选方法包括使细胞与包含位于平行位于第一和第二重组位点之间的哺乳动物细胞中起作用的复制起点的载体接触。 另一个优选的方法部分地包括使细胞与包含反平行取向的第一和第二重组位点的载体接触,使得载体被细胞内化。 在两种方法中,细胞还提供了一种位点特异性重组酶,其实现载体的第一和第二重组位点之间的重组。
    • 7. 发明申请
    • METHOD OF ANALYZING BINDING INTERACTIONS
    • 分析相互作用的方法
    • US20120077691A1
    • 2012-03-29
    • US13236651
    • 2011-09-20
    • Robert DuBridge
    • Robert DuBridge
    • C40B30/04
    • C12N15/1037C07K16/005C07K16/22C40B20/02
    • The invention is directed to methods for obtaining statistically significant information about how structural elements of proteins, e.g. position and identity of amino acid residues in binding domains, relate to functional properties of interest, such as binding affinity, specificity, and the like. In some embodiments, such information is collected by reacting under binding conditions a focused library of candidate nucleic acid-encoded binding compounds with a ligand, so that complexes form between the ligand and a portion of the candidate binding compounds (“binders”). Samples of binders and non-binders arc then decoded by high throughput nucleic acid sequencing to give statistically significant data about the binding properties of substantially all of the candidate binding compounds, permitting them to be ranked by their respective affinities or dissociation constants. A reference compound, such as a pre-existing antibody, may be included in the reaction to identify candidates with similar or improved binding characteristics that have additional desirable characteristics, such as higher solubility, reduced immunogenicity, higher stability, or the like.
    • 本发明涉及用于获得关于蛋白质的结构元件的统计学显着信息的方法,例如, 结合域中氨基酸残基的位置和特征与感兴趣的功能性质有关,例如结合亲和力,特异性等。 在一些实施方案中,通过在结合条件下使候选核酸编码的结合化合物的集中文库与配体反应来收集这样的信息,使得配体与候选结合化合物(“结合物”)的一部分之间形成复合物。 粘合剂和非粘合剂的样品然后通过高通量核酸测序解码,以得到关于基本上所有候选结合化合物的结合特性的统计学显着数据,允许它们通过它们各自的亲和力或解离常数进行排序。 可以在反应中包括参考化合物,例如预先存在的抗体,以鉴定具有相似或改善的结合特征的候选物,其具有额外的期望特征,例如更高的溶解度,降低的免疫原性,更高的稳定性等。
    • 8. 发明申请
    • MULTI-DIMENSIONAL SELECTION OF PROTEIN MUTANTS USING HIGH THROUGHPUT SEQUENCE ANALYSIS
    • 使用高通量序列分析进行多维选择蛋白质突变体
    • US20120258866A1
    • 2012-10-11
    • US13424383
    • 2012-03-20
    • Robert DuBridge
    • Robert DuBridge
    • C40B20/00
    • C07K16/005C07K16/22C07K2317/24C07K2317/565C07K2317/92C12N15/1034C12N15/1037C40B60/04
    • The invention is directed to methods for simultaneously improving a plurality of characteristics of a protein binding compound. In accordance with one aspect of the invention, a focused library of nucleic acid-encoded variants is produced and separately exposed to a plurality of reaction conditions each designed to segregate the library variants according to a different characteristic of interest, such as affinity, stability, cross-reactivity, or the like. In various embodiments, such reactions may be conducted pair-wise to simultaneously obtain improvements in two characteristics or they may be conducted three-at-a-time to simultaneously obtain improvements in three characteristics. In each case, nucleotide sequences encoding library variants segregated into improved subsets are determined, after which sequences occurring in two or more subsets are identified to obtain library variants with two or more improved characteristics.
    • 本发明涉及同时改善蛋白质结合化合物的多种特征的方法。 根据本发明的一个方面,产生核酸编码变体的集中文库,并分别暴露于多个反应条件,每个反应条件被设计为根据不同的目标特征分离文库变体,例如亲和力,稳定性, 交叉反应性等。 在各种实施方案中,这些反应可以成对进行以同时获得两个特征的改进,或者它们可以一次三次进行,以同时获得三个特征的改进。 在每种情况下,确定分离成改进子集的编码文库变体的核苷酸序列,之后鉴定出两个或多个子集中出现的序列以获得具有两个或更多个改进特征的文库变体。