Rabbit Monoclonal Antibodies: A Comparative Study Between a Novel Category of Immunoreagents and the Corresponding Mouse Monoclonal Antibodies

Abstract

Rabbit monoclonal antibodies (RabMAbs) represent a novel category of immunoreagents that may combine the best properties of both mouse monoclonal antibodies (MMAs) and of rabbit antisera. In the attempt to verify the performance of this new class of antibodies on paraffin-embedded tissue, RabMAbs against estrogen receptor, progesterone receptor, Ki-67, cyclin D1, CD3, CD5, CD23, and synaptophysin were tested on several tumor types as well as normal tissues. The results were compared with those obtained with classic MMAs against the same antigens. RabMAbs appear to offer increased sensitivity with no apparent loss of specificity. On routine use they permit higher working dilutions (5 to 10 times on average), allowing significant improvement in terms of laboratory efficiency. The robustness of RabMAbs is further proved by the fact that in some instances optimal staining can be obtained even without antigen retrieval. In consideration of the high performance observed, routine use of RabMAbs may contribute significantly to standardize diagnostic immunohistochemical procedures.

A comparison between the novel rabbit monoclonal antibodies (SP1 and B644) and mouse antibodies for evaluating estrogen receptor in breast tumors

“…There are few other studies in literature comparing the novel rabbit monoclonal antibodies to mouse antibodies (3,4,24) and no studies comparing both rabbit antibodies. Cano et al (3) evaluated ER and PR in fine-needle aspirates and paraffin-embedded sections from breast cancers using SP1 and SP2 rabbit antibodies.…”

“…The technology to prepare these antibodies from a single hybridoma allows the production of antibodies with high sensitivity and specificity, high working dilutions and better cost versus benefits. According to the suppliers, the novel rabbit monoclonal antibodies could replace the mouse monoclonal antibodies because of their high specificity, lower cost, and faster technical procedure, which would not require an antigen retrieval step (3,14,24) . Several anti-ER antibodies have been used in clinical and research immunohistochemical testing, including mouse monoclonal antibodies.…”

“…Clones 6F11 and 1D5 have been compared in clinical studies and were shown to have similar sensitivities (28) . Studies have also compared 1D5 with a novel rabbit monoclonal, SP1 (15,24) , which has eight-fold higher affinity (15) . Another study showed that SP1 is more sensitive than 1D5 for detecting ER expression in breast cancer, in both a duplicate-redundancy 431-sample TMA, and in 121 whole sections of clinical materials from multiple institutions (27) .…”

Multiple Immunoenzyme Staining: Methods and Visualizations for the Observation With Spectral Imaging

“…In the fall of 2007, the MultiVision kit system for staining a rabbit-mouse primary antibody combination including red and blue chromogens was introduced by Thermo Scientific/ LabVision. Many double staining combinations can be composed of any mouse monoclonal antibody combined with either a classical rabbit polyclonal antibody or one of the recently introduced commercially available rabbit monoclonal antibodies (Rossi et al 2005). A biotin-free detection system is applied using single species anti-mouse and anti-rabbit polymers attached with either HRP or AP enzymes (Sabattini et al 1998).…”

Predictive factors of breast cancer evaluated by immunohistochemistry

“…Recently, a new generation of rabbit monoclonal antibodies has been developed and is commercially available, such as clones SP1 and B644 anti-ER and SP2 and B645 anti-PR (24,25,46) . The technology to prepare these antibodies from a single hybridoma allows the production of antibodies with high sensitivity and specificity, high working dilutions and better cost versus benefits.…”