rev: November 11, 1999

HOME (index page)

Return  (alphabetical antibody page)

Return  (anti-Rat/Mouse Immunoglobulin..index)

  anti-RAT and MOUSE ANTIBODIES  

Background Information

RDI Division of Fitzgerald Industries Intl  offers a wide line of  antibodies. Since no one antibody works best for all applications (neutralization, blotting, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below.

Some Words on Rat Monoclonal Hybridomas

The first model of monoclonal antibody (MAb) secreting hybridomas developed by Köhler and Milstein1, was the mouse model. It can almost be considered perfect. Its efficiency and extreme stability probably account for its success. However, there are several reasons for developing another rodent model, particularly a rat model.

The first reason is certainly linked to the rat antibody repertoire which is different from that of the mouse. This is true for the mouse xenoantigens which can be studied with rat but not with mouse MAbs. Likewise, rat alloantigens can be better studied with rat than with mouse MAbs. At last, some antigens from origins other than mouse or rat species can induce much stronger immune responses in rats than in mice and evidently the reciprocal could be true for other antigens.

The second reason relates to physicochemical and biological properties of rat immunoglobulins which do not seem to exist in the mouse species. Rat antibodies of the IgG1, IgG2a, and IgG2b isotypes can easily fix the human or the rabbit complement. Likewise, the rat IgG2b antibodies can be efficiently used by human K cells to kill their target cells.

The third reason is linked to the in vivo production of rat MAbs. Rats are approximately ten times bigger than mice. A LOU/C rat could give a mean production of 50 to 150 mg of MAbs per animal. Although the future of the production of MAbs lies in in vitro culture, at the present time, MAbs in vivo production is less animal cosuming in rats than in mice.

The fourth reason concerns the purification methods which have been set up for rat MAbs; they are easy and rapid to use, highly efficient, and inexpensive. An in vitro culture supernatant2 and an in vivo ascitic fluid method3 have been developed for purification of rat MAbs by immunoaffinity chromatography. However, conventional purification techniques of rat immunoglobulins can also be used, but these techniques are based on properties of heterogenous molecule populations which are rarely found in a given MAb preparation. Techniques of DEAE chromatography, gel filtration, or preparative electrophoresis can be used, but the degree of purity and the percentage of recovery achieved by them are limited. In most
cases, affinity chromatography is preferable.

The fith reason is the absence of viral particles in rat hybridomas and therefore in rat MAbs; this is an important fact to take into account in the case of human in vivo therapeutic use.

However, obtaining rat-rat hybridomas still seems to be considered difficult. In sentences like, "At the time of writing, successful production of rat-rat hybridomas have been limited to a small number of laboratories. Several experienced investigators have had difficulty in keeping rat-rat hybrids alive for more than 2 to 3 weeks ..."4, the same author in the second edition of his book5 adds to the same paragraph, "The reason for such problems is not yet clear. The hybrids seem to grow initially, and then die. Perhaps rat cells are unduly susceptible to 'natural killer' cells, or some rat colonies have unusually high number of natural killer cells in the spleen". Similar sentences are frequently written or heard in conferences and seminars. Already we have obtained, and so have scientists around us, thousands and thousands of rat-rat hybridomas with no more difficulty than if we are making mouse-mouse hybridomas. We do not believe that we are especially clever, but rather we believe it is just a question of experiments which can be communicated to interested colleagues.

Moreover, many scientists are not familiar with rat immunoglobulins and rat MAbs. There are no books and no reviews devoted either to the development of rat hybridomas or to the use of rat MAbs. The purpose of "Rat Hybridomas and Rat Monoclonal antibodies" H. BAZIN (Ed.) CRC Press, Boca Raton, Florida, 1990, 515 pages is to fill this gap. However, it should be made clear that we have not tried by any means to write an exhaustive review on hybridomas in general as many excellent books have already been written on this subject, although they generally concern mouse4-7 or human hybridomas8,9 which are now too numerous to all be described or on rat hybridomas. We have attempted rather to write on our own experiments or on those of friend laboratories which have used similar techniques against many different kinds of antigens.

We hope that this book will further the development of the use of the rat hybridomas and the rat monoclonal antibodies.

REFERENCES

1. Köhler G. and Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, 256:           495, 1975.

2. Bazin H., Xhurdebise L.M., Burtonboy G., Lebacq A.M., De Clercq L., and Cormont F. Rat monoclonal                    
    antibodies.I.Rapid purification from in vitro culture supernatants. J. Immunol. Methods, 66: 261, 1984.

3. Bazin H., Cormont F., and De Clercq L. Rat monoclonal antibodies.II. Rapid and efficient method of purification from
    ascitic fluid or serum. J. Immunol. Methods, 71, 9: 1984.

4. Goding J.W. Monoclonal Antibodies: Principle and Practice, 1st ed., Academic Press, London, 1983, 276.

5. Goding J.W. Monoclonal Antibodies: Principle and Practice, 2nd ed., Academic Press, London, 1986, 315.

6. Hurrell J.G.R. Monoclonal Hybridoma Antibodies: Techniques and Applications, CRC Press, Boca Raton, FL, 1985,
    240.

7. Bartal H., and Hirshaut Y. Methods of Hybridoma Formation. Humana Press, Clifton, NJ, 1987.

8. Engleman E.G., Foung S.K.H., Larrick J., and Raubitschek A. Human Hybridomas and Monoclonal Antibodies. Plenum
    Press, New York, 1985.

9. Strelkauskas A.J. Human Hybridomas - Diagnosis and Therapeutic Applications, Marcel Dekker, Inc., New York,
    1987.

Vials

1 or 5 or 10 or more ml.

Concentration

1 mg/ml or other concentrations upon request.

Purified products

They are in PBS buffer + NaN3 0.1%

Some purified LO-CD (in order to be used in cell cultures) can be in PBS buffer without NaN3 upon request

Labelled products

- With Biotin and with FITC: are in PBS buffer + 0.1% NaN3 and glycerol 50%

- With Peroxidase : are in PBS buffer + glycerol 50%

Ascitic fluid

Preservative reagents:

- PMSF (phenyl methane sulfonyl fluorid): 20 µl/ml of ascite from a solution at 40 mg/ml in ethanol.

- Thimerosal: 10 µl/ml of ascitic fluid from a solution at 1g/l of distilled water.

Storage of the products:

They must be stored at -20°C.

The nomenclature of the Rat Immunoglobulin Isotypes has been first described in:

Bazin H., Beckers A., Querinjean P.Three classes and four (sub)classes of rat immunoglobulins : IgM, IgA, IgE and IgG1, IgG2a, IgG2b, IgG2c.European Journal of Immunology, 1974, 4 : 44-48.

Bazin H., Beckers A., Urbain-Vansanten G., Pauwels R., Bruyns C., Tilkin A.F., Platteau B., Urbain J.Transplantable IgD immunoglobulin-secreting tumours in rats.Journal of Immunology, 1978, 121 : 2077-2082.

The ICDB numbers correspond to the numbers attributed by the Immunoclone Database Bank to "Hybridoma-monoclonal antibody".

RDI Division of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

Return  (anti-Rat/Mouse Immunoglobulin..index)

Ordering terms