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Rabbit vs. Mouse Monoclonal Antibodies

Rabbit vs. Mouse Monoclonal Antibodies

Epitomics has developed a unique and proprietary method for making monoclonal antibodies from rabbits rather than the conventional method of starting with mice. The chart below demonstrates the general outline that Epitomics uses for producing a rabbit monoclonal antibody.

The basic principal for making the antibody is the same as for mouse monoclonals. Our proprietary Rabbit fusion partner cells can fuse to rabbit B-cells to create the Rabbit Hybridoma cells. Hybridomas are then screened to select for clones with specific and sensitive antigen recognition and the antibodies are characterized using a variety of methods.

Advantages of Rabbit Monoclonals

More diverse epitope recognition
Improved immune response to small-size epitopes
High specificity and affinity
Greatly improved response to mouse antigens

To put it simply - better antigen recognition. The rabbit immune system generates antibody diversity and optimizes affinity by mechanisms that are more efficient than those of mice and other rodents. This increases the possibility of obtaining a functional antibody that will work in a variety of applications. Additionally, many small compounds and peptides do not elicit a good immune response in mice but do so in rabbits. It is often for this reason, that rabbit polyclonal antibodies are developed and used in many research and even some diagnostic applications, ranging from drug screening to clinical diagnosis. As an example of this, most of the phospho specific recognition antibodies being sold on the market today are rabbit polyclonals.

Additionally, many small compounds and peptides do not elicit a good immune response in mice but do so in rabbits. It is often for this reason, that rabbit polyclonal antibodies are developed and used in many research and even some diagnostic applications, ranging from drug screening to clinical diagnosis. As an example of this, most of the phospho specific recognition antibodies being sold on the market today are rabbit polyclonals.

Comparison of Rabbit and Mouse Monoclonal Antibodies

	Mouse	Rabbit
Antigen recognition:	• Limited immuno response	• High chance of success with range of antigens including small molecules and peptides
	• Small molecules/peptides often non-immunogenic	• Good response to use rodent proteins
	• Very limited response to rodent antigens	• Recognizes several epitopes per protein antigen
	• Recognizes limited number of epitopes (due to immunodominance)
Affinities:	Nanomolar (~10-9 KD M)	Picomolar (10-12 KD M) possible
Specificity:	Med-High	High
Applications:	Westerns, ELISA, Flow Cytometry, IP, not always suitable for IHC, ICC.	Westerns, ELISA, Flow Cytometry, IP, IHC, ICC (excellent results in IHC).

See below for IHC Comparison images - on Formalin fixed, paraffin-embedded Human tissue

CDX2

Name	Catalog#	Applications
CDX2	2475-1	WB, UHC, ICC, FC

Paraffin-embedded human colonic carcinoma tissue stained with Epitomics' CDX2 RabMAb (Cat# 2475-1) and Vendor A's CDX2 Mouse Monoclonal (both at 1:1000 dilution factor)

Cathepsin D

Name	Catalog#	Applications
CathespinD	2487-1	WB, IHC, ICC
CathespinD	2431-1	WB, IHC, FC, IP
CathespinD pro	2510-1	WB

Paraffin-embedded human breast ductal infiltrating carcinoma tissue stained with Epitomics' Cathespsin D RabMAb (Cat# 2510-1, at 1:500 dilution factor) and Vendor A's Cathespin D Mouse Monoclonal (at 1:50 dilution factor)

E-Cadherin

Name	Catalog #	Applications
E-Cadherin	1702-1	WB, IHC, ICC, IP
E-Cadherin (pS838/840)	2239-1	WB, IHC, ICC, IP

Paraffin-embedded human breast carcinoma tissue stained with Epitomics' E-Cadherin RabMAb (Cat# 1702-1, at 1:500 dilution factor) and Vendor A's E-Cadherin Mouse Monoclonal (at 1:50 dilution factor).

Her2

Name	Catalog #	Applications
Her2	2064-1	WB, IHC, FC, IP
Her2	4201-1	IHC
Her2 (pY877)	2265-1	WB, ICC
Her2 (p1139)	1191-1	WB, FC, IP

Paraffin-embedded human breast carcinoma tissue stained with Epitomics' Her2 RabMAb and Vendor A's Her2 Mouse Monoclonal (both at 1:500 dilution factor).

MCM5

Name	Catalog #	Applications
MCM5	2380-1	WB, IHC, ICC, FC, IP
MCM5	2374-1	WB, IHC, ICC, FC, IP

Paraffin-embedded human brain glioma tissue stained with Epitomics' MCM5 RabMAb (Cat# 2374-1) and Vendor A's MCM5 Mouse Monoclonal (both at 1:500 dilution factor)

Rabbit vs. Mouse References

Rabbit monoclonal antibodies show higher sensitivity than mouse monoclonals for estrogen and progesterone receptor evaluation in breast cancer by immunohistochemistry. Rocha R, Nunes C, Rocha G, Oliveira F, Sanches F, Gobbi H. [Pathol Res Pract, 2008;204(9):655-62. Epub 2008 Jun 18]

Comparative analysis of six different antibodies against Her2 including the novel rabbit monoclonal antibody (SP3) and chromogenic in situ hybridisation in breast carcinomas. Nunes CB, Rocha RM, Reis-Filho JS, Lambros MB, Rocha GF, Sanches FS, Oliveira FN, Gobbi H. [J Clin Pathol. 2008 Aug;61(8):934-8. Epub 2008 May 12]

Rabbit monoclonal antibodies: a comparative study between a novel category of immunoreagents and the corresponding mouse monoclonal antibodies. Rossi S, Laurino L, Furlanetto A, Chinellato S, Orvieto E, Canal F, Facchetti F, Dei Tos AP. [Am J Clin Pathol. 2005 Aug;124(2):295-302]

A comparison between the novel rabbit monoclonal antibodies (SP1 and B644) and mouse antibodies for evaluating estrogen receptor in breast tumors. Rafael Malagoli Rocha1; Cristiana Buzelin Nunes, Gislene Fátima Silva Rocha, Flávio Nepomuceno Oliveira, Fernanda Squárcio Fernandes Sanches, Helenice Gobbi [J Bras Patol Med Lab. 2007;143(6):435-440]

A new rabbit monoclonal antibody (4B5) for the immunohistochemical (IHC) determination of the HER2 status in breast cancer: comparison with CB11, fluorescence in situ hybridization (FISH), and interlaboratory reproducibility. Powell WC, Hicks DG, Prescott N, Tarr SM, Laniauskas S, Williams T, Short S, Pettay J, Nagle RB, Dabbs DJ, Scott KM, Brown RW, Grogan T, Roche PC, Tubbs RR. [Appl Immunohistochem Mol Morphol. 2007 Mar;15(1):94-102]

Immunohistochemical Detection Using the New Rabbit Monoclonal Antibody SP1 of Estrogen Receptor in Breast Cancer Is Superior to Mouse Monoclonal Antibody 1D5 in Predicting Survival. Maggie C.U. Cheang, Diana O. Treaba, Caroline H. Speers, Ivo A. Olivotto, Chris D. Bajdik, Stephen K. Chia, Lynn C. Goldstein, Karen A. Gelmon, David Huntsman, C. Blake Gilks, Torsten O. Nielsen, Allen M. Gown [Journal of Clinical Oncology, Vol 24, No 36 (December 20), 2006: pp. 5637-5644]

To evaluate the specificity and sensitivity of Epitomics' RabMAbs, we compared various diagnostic marker specific RabMAbs with the leading commercially available Mouse Monoclonal counterparts. Both antibodies were tested on the same formalin fixed paraffin-embedded tissue sample and under their recommended immunohistochemistry conditions.

The results shown below demonstrate that Epitomics' RabMAbs have higher sensitivity combined with lower background than the comparable mouse monoclonal antibodies. In conclusion, Epitomics RabMAbs are proving to be exceptional affinity reagents for the detection of key diagnostic and prognostic markers in formalin fixed and paraffin embedded (FFPE) tissue sample.

Patented Technology

Epitomics RabMab technology is the only currently available hybridoma based technology for making rabbit monoclonals and covered under the following U.S. Patents, No. 5,675,063 and 7,429,487. Our broad patents cover not only the rabbit fusion partner cell line but also the method for generating rabbit fusion partner cell line as well as the antibodies produced from the cell line.

See IHC Comparison images

Rabbit Antibodies Sample Size

For you to test the Rabbit monoclonal antibodies, all Epitomics products are now available in 40ul vials. These sample packages are just € 75 a piece and give you a chance to see the advantages of rabbit monoclonal antibodies for yourself.