Monoclonal Antibodies against Human β‐Glucocerebrosidase

Barneveld, R. A.; Tegelaers, F. P. W.; Ginns, Edward I.; Visser, Pim; Laanen, Elly A.; Brady, Roscoe O.; Galjaard, H.; Barranger, John A.; Reuser, Arnold; Tager, J. M.

doi:10.1111/j.1432-1033.1983.tb07606.x

Cited by 46 publications

(18 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Studies now completed in four collaborating laboratories, using a highly specific, high-titre rabbit antiserum and monoclonal antibodies raised against homogeneous glucocerebrosidase, have unequivocally demonstrated multiple processing forms of glucocerebrosidase in control fibroblasts [18 -22, 341. Several other observations confirm the identity of the species immunoprecipitated in these studies. The most compelling evidence is that not only the polyclonal rabbit antiserum but also the monoclonal antibody 8E4 [35] is able to produce the three bands of cross-reactive material characteristic of normal fibroblast glucocerebrosidase [18,19]. In addition, both the antiserum and the monoclonal antibody are capable of immunoprecipitating about 95% of the glucocerebrosidase activity in fibroblast extracts These observations, taken together, prove conclusively that the species under study is indeed glucocerebrosidase and not some contaminant protein of similar molecular mass.…”

Section: Discussionmentioning

confidence: 99%

Biosynthesis and maturation of glucocerebrosidase in Gaucher fibroblasts

et al. 1987

Self Cite

View full text Add to dashboard Cite

The biosynthesis and maturation of glucocerebrosidase were studied in fibroblasts from patients with the neurological and non-neurological forms of Gaucher disease and in control cells. In control fibroblasts the precursor of glucocerebrosidase (62 -63 kDa), observed after a short pulse with [35S]methionine, was converted during the chase period to a 66-kDa intermediate form and, finally, to the 59-kDa mature protein. In fibroblasts from patients with the non-neurological phenotype of Gaucher disease (type 1) the same biosynthetic forms were seen as in control fibroblasts. These biosynthetic forms correspond to the three-banded pattern seen in control and Gaucher type 1 fibroblast extracts analysed by the immunoblotting procedure, or after electrophoresis and fluorography of extracts of such fibroblasts cultured for 5 days with [14C]leucine. The 59-kDa protein seen in type 1 fibroblasts was unstable and disappeared after a prolonged chase; this disappearance was not observed when the cells were grown in the presence of leupeptin. In fibroblasts from patients with the neurological forms of Gaucher disease (types 2 and 3) the 62.5-kDa precursor of glucocerebrosidase was present in near-normal amounts after a short pulse, but the 59-kDa form was not detected even when cells were cultured with leupeptin.

show abstract

Section: Discussionmentioning

confidence: 99%

Biosynthesis and maturation of glucocerebrosidase in Gaucher fibroblasts

et al. 1987

Self Cite

View full text Add to dashboard Cite

show abstract

“…The MAbs 2C7 and A2C6, used as control antibodies, are also of the IgGI isotype. MAb 2C7 reacts with human a-glucocerebrosidase (Barneveld et al, 1983) and MAb A2C6 reacts with the hepatitis B surface antigen (Zurawski et al, 1983). Purified IgG of MAbs OC125 and OV-TL 3 was kindly provided by Dr S.O.…”

Section: Methodsmentioning

confidence: 99%

Comparison of the pharmacokinetics, biodistribution and dosimetry of monoclonal antibodies OC125, OV-TL 3, and 139H2 as IgG and F(ab')2 fragments in experimental ovarian cancer

Molthoff

Pinedo²,

Schlüper³

et al. 1992

Br J Cancer

View full text Add to dashboard Cite

Monoclonal antibody (MAb) 139H2 was previously shown to localise specifically into ovarian cancer xenografts in nude mice. MAb 139H2 was compared with MAbs OC125 and OV-TL 3, all reactive with ovarian carcinomas, for the binding characteristics as IgG and F(ab')2 fragments with the use of the OVCAR-3 cell line grown in vitro and as s.c. xenografts. Immunoperoxidase staining of OVCAR-3 tissue sections with MAbs OC125 and 139H2 was heterogeneous, whereas MAb OV-TL 3 showed homogeneity. No differences in binding were observed between IgG and F(ab')2. The avidity expressed as apparent affinity constants of MAbs OC125, OV-TL 3 and 139H2 for OVAR-3 cells were 1 x 10(9) M-1, 1 x 10(9) M-1, and 1 x 10(8) M-1, while the number of antigenic determinants were 5 x 10(6), 1 x 10(6) and 7 x 10(6), respectively. In OVCAR-3 bearing nude mice the blood half-lives of the MAbs as IgG and F(ab')2 were approximately 50 h and 6 h, respectively. Maximum tumour uptake for the whole MAbs OC125, OV-TL 3, 139H2 and a control MAb 2C7 was 8.5%, 17.7%, 11.1% and 2.5% of the injected dose g-1, reached at 72 h after injection. For the respective F(ab')2 fragments, the maximum values were 5.2%, 10.0%, 5.5% and 1.9% of the injected dose g-1, reached between 6 h and 15 h. Tumour to non-tumour ratios were more favourable for the F(ab')2 fragments as compared to those for MAbs as IgG. Biodistribution in mice bearing a control tumour confirmed the specificity of tumour localisation of MAbs OC125, OV-TL 3 and 139H2. After injection of a tracer dose of 10 microCi of radiolabelled MAbs OC125, OV-TL 3 and 139H2 as IgG, tumours received 38 cGy, and 9 cGy. In our OVCAR-3 model, a ranking in efficiency in tumour localisation would indicate MAb OV-TL 3 as most favourable MAb, but cross-reactivity with subpopulations of human white blood cells might hamper its clinical use. Dosimetric data indicate a 4-fold higher radiation absorbed dose to tumours for IgG compared with F(ab')2 fragments.

show abstract

“…Immunochemical detection was performed as described by van Dongen et al (28). The mouse monoclonal anti-human GC antibody 8E4 was used to detect human GC (29). Fluorescein-conjugated goat anti-mouse immunoglobins (Vector Laboratories) were used as secondary antibodies.…”

Section: Methodsmentioning

confidence: 99%

Production of human glucocerebrosidase in mice after retroviral gene transfer into multipotential hematopoietic progenitor cells.

Correll

Fink

Brady

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The human glucocerebrosidase (GC) gene has been transferred efficiently into spleen colony-forming unit (CFU-S) multipotential hematopoietic progenitor cells, and production of human GC RNA and protein has been achieved in transduced CFU-S colonies. High-titer retroviral vectors containing the human GC cDNA were constructed. Mouse bone marrow cells were stimulated with hematopoietic growth factors, infected by coculture with producer cells, and injected into lethally irradiated animals. Four vectors were compared with respect to gene-transfer efficiency into CFU-S progenitors. One vector (G vector) required high concentrations of interleukins 3 and 6 during stimulation and coculture for efficient transduction of CFU-S progenitors. The remaining three vectors (NTG, GTN, and GI vectors) transduced these progenitors at infection frequencies approaching 100% using low concentrations of hematopoietic growth factors to stimulate cell division prior to and during the infection. Vectors using the viral long terminal repeat enhancer/promoter to drive the human GC cDNA produced high levels of human GC RNA in the progeny of CFU-S progenitors after gene transfer. When an internal herpes simplex thymidine kinase promoter assisted by a mutant polyoma enhancer was used to drive the human GC cDNA (NTG vector), little or no human GC RNA was detected in transduced CFU-S colonies. All three vectors producing human GC RNA in CFU-S colonies can generate human GC as detected by immunochemical analysis of CFU-S colonies. NTG vector-infected bone marrow cells were transplanted into W/Wv recipients to generate long-term reconstituted mice. The capacity of the viral long terminal repeat and the internal thymidine kinase promoter to direct synthesis of RNA in transduced bone marrow and spleen cells 5 months after bone marrow transplantation reflected the performance of these promoters in NTG-transduced CFU-S colonies.

show abstract

Monoclonal Antibodies against Human β‐Glucocerebrosidase

Cited by 46 publications

References 21 publications

Biosynthesis and maturation of glucocerebrosidase in Gaucher fibroblasts

Biosynthesis and maturation of glucocerebrosidase in Gaucher fibroblasts

Comparison of the pharmacokinetics, biodistribution and dosimetry of monoclonal antibodies OC125, OV-TL 3, and 139H2 as IgG and F(ab')2 fragments in experimental ovarian cancer

Production of human glucocerebrosidase in mice after retroviral gene transfer into multipotential hematopoietic progenitor cells.

Contact Info

Product

Resources

About