The present invention provides labeling reagents and methods for labeling primary antibodies and for detecting a target in a sample using an immuno-labeled complex that comprises a target-binding antibody and one or more labeling reagents. The labeling reagents comprise monovalent antibody fragments or non-antibody monomeric proteins whereby the labeling reagents have affinity for a specific region of the target-binding antibody and are covalently attached to a label. Typically, the labeling reagent is an anti-Fc Fab or Fab′ fragment that was generated by immunizing a goat or rabbit with the Fc fragment of an antibody. The present invention provides for discrete subsets of labeling reagent and immuno-labeled complexes that facilitate the simultaneous detection of multiple targets in a sample wherein the immuno-labeled complexes are distinguished by i) a ratio of label to labeling reagent, or ii) a physical property of said label, or iii) a ratio of labeling reagent to said target-binding antibody, or iv) by said target-binding antibody. This is particularly useful for fluorophore labels that can be attached to labeling reagents and subsequently immuno-labeled complexes in ratios for the detection of multiple targets.
本发明提供了标记试剂和标记原初
抗体的方法,以及使用包含靶向结合
抗体和一种或多种标记试剂的免疫标记复合物在样品中检测目标的方法。标记试剂包括单价
抗体片段或非
抗体单体蛋白,其中标记试剂具有亲和力,能够与靶向结合
抗体的特定区域结合,并与标记共价连接。通常,标记试剂是通过用兔子或山羊免疫
抗体的Fc片段生成的抗Fc Fab或Fab'片段。本发明提供了离散的子集标记试剂和免疫标记复合物,有助于在样品中同时检测多个目标,其中免疫标记复合物通过i)标记与标记试剂的比率,或ii)标记的物理特性,或iii)标记试剂与靶向结合
抗体的比率,或iv)靶向结合
抗体来区分。这对于荧光标记特别有用,可以将其附加到标记试剂和随后的免疫标记复合物中,以检测多个目标。