Pilot scale purification of human monoclonal IgM (COU-1) for clinical trials

Ida Tornøe, Ingrid L. Titlestad*, Karin Kejling, Karin Erb, Henrik J. Ditzel, Jens C. Jensenius

*Kontaktforfatter for dette arbejde

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

No standard procedure is available for the purification of human monoclonal antibodies for human i.v. administration. Here we describe the procedure developed for pilot scale purification of the human IgM monoclonal antibody COU-1 directed against a cancer-associated antigen. The hybridoma cells were grown in protein-free medium and purification from the clarified culture supernatant was carried out in 4 simple chromatographic steps: (1) hydroxylapatite chromatography: (2) hydrophobic interaction chromatography on phenyl-Sepharose: (3) cation-exchange chromatography on sulphonyl-Sepharose; and (4) anion-exchange chromatography on tetraethylamino-Sepharose. The product was substantially pure with regard to protein after step 3, but contained DNA which was removed in step 4. The average recovery of the IgM was 54% with a range of 40-65%. Importantly, the ability of the antibody to bind to its antigen in ELISA was fully maintained during the purification. Subsequently, the purified antibody was isotope labelled and successfully used for in vivo detection of colon, rectal and pancreas carcinomas in patients. The purification procedure described appears to compare favourably with previously published methods, but a critical comparison is not possible due to the lack of necessary information in the available literature.

OriginalsprogEngelsk
TidsskriftJournal of Immunological Methods
Vol/bind205
Udgave nummer1
Sider (fra-til)11-17
Antal sider7
ISSN0022-1759
DOI
StatusUdgivet - 23. jun. 1997

Fingeraftryk

Chromatography
Clinical Trials
Sepharose
Serum-Free Culture Media
Hybridomas
Durapatite
Isotopes
Cations
Pancreas
Colon
DNA
Neoplasms
Proteins

Citer dette

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title = "Pilot scale purification of human monoclonal IgM (COU-1) for clinical trials",
abstract = "No standard procedure is available for the purification of human monoclonal antibodies for human i.v. administration. Here we describe the procedure developed for pilot scale purification of the human IgM monoclonal antibody COU-1 directed against a cancer-associated antigen. The hybridoma cells were grown in protein-free medium and purification from the clarified culture supernatant was carried out in 4 simple chromatographic steps: (1) hydroxylapatite chromatography: (2) hydrophobic interaction chromatography on phenyl-Sepharose: (3) cation-exchange chromatography on sulphonyl-Sepharose; and (4) anion-exchange chromatography on tetraethylamino-Sepharose. The product was substantially pure with regard to protein after step 3, but contained DNA which was removed in step 4. The average recovery of the IgM was 54{\%} with a range of 40-65{\%}. Importantly, the ability of the antibody to bind to its antigen in ELISA was fully maintained during the purification. Subsequently, the purified antibody was isotope labelled and successfully used for in vivo detection of colon, rectal and pancreas carcinomas in patients. The purification procedure described appears to compare favourably with previously published methods, but a critical comparison is not possible due to the lack of necessary information in the available literature.",
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author = "Ida Torn{\o}e and Titlestad, {Ingrid L.} and Karin Kejling and Karin Erb and Ditzel, {Henrik J.} and Jensenius, {Jens C.}",
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Pilot scale purification of human monoclonal IgM (COU-1) for clinical trials. / Tornøe, Ida; Titlestad, Ingrid L.; Kejling, Karin; Erb, Karin; Ditzel, Henrik J.; Jensenius, Jens C.

I: Journal of Immunological Methods, Bind 205, Nr. 1, 23.06.1997, s. 11-17.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Pilot scale purification of human monoclonal IgM (COU-1) for clinical trials

AU - Tornøe, Ida

AU - Titlestad, Ingrid L.

AU - Kejling, Karin

AU - Erb, Karin

AU - Ditzel, Henrik J.

AU - Jensenius, Jens C.

PY - 1997/6/23

Y1 - 1997/6/23

N2 - No standard procedure is available for the purification of human monoclonal antibodies for human i.v. administration. Here we describe the procedure developed for pilot scale purification of the human IgM monoclonal antibody COU-1 directed against a cancer-associated antigen. The hybridoma cells were grown in protein-free medium and purification from the clarified culture supernatant was carried out in 4 simple chromatographic steps: (1) hydroxylapatite chromatography: (2) hydrophobic interaction chromatography on phenyl-Sepharose: (3) cation-exchange chromatography on sulphonyl-Sepharose; and (4) anion-exchange chromatography on tetraethylamino-Sepharose. The product was substantially pure with regard to protein after step 3, but contained DNA which was removed in step 4. The average recovery of the IgM was 54% with a range of 40-65%. Importantly, the ability of the antibody to bind to its antigen in ELISA was fully maintained during the purification. Subsequently, the purified antibody was isotope labelled and successfully used for in vivo detection of colon, rectal and pancreas carcinomas in patients. The purification procedure described appears to compare favourably with previously published methods, but a critical comparison is not possible due to the lack of necessary information in the available literature.

AB - No standard procedure is available for the purification of human monoclonal antibodies for human i.v. administration. Here we describe the procedure developed for pilot scale purification of the human IgM monoclonal antibody COU-1 directed against a cancer-associated antigen. The hybridoma cells were grown in protein-free medium and purification from the clarified culture supernatant was carried out in 4 simple chromatographic steps: (1) hydroxylapatite chromatography: (2) hydrophobic interaction chromatography on phenyl-Sepharose: (3) cation-exchange chromatography on sulphonyl-Sepharose; and (4) anion-exchange chromatography on tetraethylamino-Sepharose. The product was substantially pure with regard to protein after step 3, but contained DNA which was removed in step 4. The average recovery of the IgM was 54% with a range of 40-65%. Importantly, the ability of the antibody to bind to its antigen in ELISA was fully maintained during the purification. Subsequently, the purified antibody was isotope labelled and successfully used for in vivo detection of colon, rectal and pancreas carcinomas in patients. The purification procedure described appears to compare favourably with previously published methods, but a critical comparison is not possible due to the lack of necessary information in the available literature.

KW - Human monoclonal antibody

KW - IgM purification

KW - Pilot scale

U2 - 10.1016/S0022-1759(97)00051-3

DO - 10.1016/S0022-1759(97)00051-3

M3 - Journal article

C2 - 9236910

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JO - Journal of Immunological Methods

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