46 ANTIGEN USED FOR IMMUNIZATION Unpurified extracts from ventricular heart muscle (human: pa- tients with myocarditis and dilated cardiomyopathy) METHOD OF IMMUNIZATION Immunization of three BALB/c mice. Details as in original paper. (1) PARENTAL CELL LINE USED FOR FUSION NS0 SELECTION AND CLONING PROCEDURE Method of Galfre et al. (2) In total, 1500 hybridomas were pro- duced, and 325 were screened. H1 and DH2 were further as- sessed. HEAVY AND LIGHT CHAINS OF IMMUNOGLOBULIN Heavy chains were H1, gamma1; and DH2, mu. Light chains were not determined. SPECIFICITY In original paper, MAbs were tested in immunocytochemistry on cryostat sections of human heart and suggested to be reactive against human myosin. (1) SPECIFIC ANTIGEN IDENTIFIED MAb H1 was reactive to human heart, porcine heart, and mouse heart; it was also reactive to porcine and bovine skeletal muscle and to a lesser extent to rabbit skeletal muscle. Potential epitope was IEFKKIVERRDALLV. (3) MAb DH2 was weakly reactive to human heart, porcine heart, and mouse heart, and also reactive to porcine and bovine skeletal muscle, and to a lesser extent to rabbit skeletal muscle. AVAILABILITY Tissue culture supernatant Yes ✓ No Ascitic fluid Yes No ✓ Hybridoma cells Yes No ✓ REFERENCES 1. Spry CJF and Tai P-C: Dilated cardiomyopathy and myocarditis: monoclonal antibodies to diseased heart tissues. Eur Heart Journal 1991;12:130–133. 2. Galfre G, Howes SC, Milstein C, Butcher GW, and Howard JC: Antibodies to major histocompatibility antigens pro- duced by hybrid cell lines. Nature 1977;266:550–552. 3. Nelson PN, Astley SJ, Roden D, Waldron E, Naig K, Caforio ALP, Koutedakis Y, Perera S, and Spry C: Characterization of anti-myosin monoclonal antibodies. Hybridoma 2005;24:314–318. ADDRESS CORRESPONDENCE TO: De Niet Sanbio Holland Telephone: 00 31-4132-51115 http://www.monosan.com HYBRIDOMA Volume 25, Number 1, 2006 © Mary Ann Liebert, Inc. MAbs H1 and DH2 Monoclonal Antibodies The hybridomas were originally developed and owned by Prof. C. Spry, who has since retired. Prof. Spry passed on his stocks of hybridomas, including H1 and DH2, to Mr. De Niet Sanbio.

MAbs H1 and DH2

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ANTIGEN USED FOR IMMUNIZATION

Unpurified extracts from ventricular heart muscle (human: pa-tients with myocarditis and dilated cardiomyopathy)

METHOD OF IMMUNIZATION

Immunization of three BALB/c mice. Details as in original pa-per.(1)

PARENTAL CELL LINE USED FORFUSION

NS0

SELECTION AND CLONING PROCEDURE

Method of Galfre et al.(2) In total, 1500 hybridomas were pro-duced, and 325 were screened. H1 and DH2 were further as-sessed.

HEAVY AND LIGHT CHAINS OFIMMUNOGLOBULIN

Heavy chains were H1, gamma1; and DH2, mu. Light chainswere not determined.

SPECIFICITY

In original paper, MAbs were tested in immunocytochemistryon cryostat sections of human heart and suggested to be reac-tive against human myosin.(1)

SPECIFIC ANTIGEN IDENTIFIED

MAb H1 was reactive to human heart, porcine heart, and mouseheart; it was also reactive to porcine and bovine skeletal mus-

cle and to a lesser extent to rabbit skeletal muscle. Potentialepitope was IEFKKIVERRDALLV.(3) MAb DH2 was weaklyreactive to human heart, porcine heart, and mouse heart, andalso reactive to porcine and bovine skeletal muscle, and to alesser extent to rabbit skeletal muscle.

AVAILABILITY

Tissue culture supernatant Yes ✓ NoAscitic fluid Yes No ✓

Hybridoma cells Yes No ✓

REFERENCES

1. Spry CJF and Tai P-C: Dilated cardiomyopathy and my-ocarditis: monoclonal antibodies to diseased heart tissues.Eur Heart Journal 1991;12:130–133.

2. Galfre G, Howes SC, Milstein C, Butcher GW, and HowardJC: Antibodies to major histocompatibility antigens pro-duced by hybrid cell lines. Nature 1977;266:550–552.

3. Nelson PN, Astley SJ, Roden D, Waldron E, Naig K, CaforioALP, Koutedakis Y, Perera S, and Spry C: Characterizationof anti-myosin monoclonal antibodies. Hybridoma2005;24:314–318.

ADDRESS CORRESPONDENCE TO:

De Niet SanbioHollandTelephone: 00 31-4132-51115http://www.monosan.com

HYBRIDOMAVolume 25, Number 1, 2006© Mary Ann Liebert, Inc.

MAbs H1 and DH2

Monoclonal Antibodies

The hybridomas were originally developed and owned by Prof.C. Spry, who has since retired. Prof. Spry passed on his stocksof hybridomas, including H1 and DH2, to Mr. De Niet Sanbio.