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Novel antitenascin antibody with increased tumour localisation for Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT).

De Santis R, Anastasi AM, D'Alessio V, Pelliccia A, Albertoni C, Rosi A, Leoni B, Lindstedt R, Petronzelli F, Dani M, Verdoliva A, Ippolito A, Campanile N, Manfredi V, Esposito A, Cassani G, Chinol M, Paganelli G, Carminati P - Br. J. Cancer (2003)

Bottom Line: The hybridoma clone producing the monoclonal antitenascin antibody BC4, previously used for clinical applications, was found not suitable for further development because of the production of an additional, nonfunctional light chain.ST2146 was found able to bind human tenascin at an epitope strictly related, if not identical, to the antigenic epitope of BC4.It showed, compared to BC4, higher affinity and immunoreactivity and similar selectivity by immunohistochemistry.

View Article: PubMed Central - PubMed

Affiliation: Immunology Department, Sigma Tau SpA R&D, Pomezia, Rome, Italy. rita.desantis@sigma-tau.it

ABSTRACT
The Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT) method is based on intravenous, sequential administration of a biotinylated antibody, avidin/streptavidin and (90)Y-labelled biotin. The hybridoma clone producing the monoclonal antitenascin antibody BC4, previously used for clinical applications, was found not suitable for further development because of the production of an additional, nonfunctional light chain. In order to solve this problem, the new cST2146 hybridoma clone was generated. The monoclonal antibody ST2146, produced by this hybridoma, having the same specificity as BC4 but lacking the nonfunctional light chain, was characterised. ST2146 was found able to bind human tenascin at an epitope strictly related, if not identical, to the antigenic epitope of BC4. It showed, compared to BC4, higher affinity and immunoreactivity and similar selectivity by immunohistochemistry. Biodistribution studies of biotinylated ST2146 and three other monoclonal antitenascin antibodies showed for ST2146 the highest and more specific tumour localisation in HT29-grafted nude mice. On the overall, ST2146 appears to be a good alternative to BC4 for further clinical development of PAGRIT.

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ELISA plates coated with tenascin at 5 μg ml−1 (A) or 0.5 μg ml−1 (B). BC4 peaks 1–3 were obtained by subjecting the protein A eluted BC4 to hydroxylapatite chromatography as shown in Figure 2B.
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fig4: ELISA plates coated with tenascin at 5 μg ml−1 (A) or 0.5 μg ml−1 (B). BC4 peaks 1–3 were obtained by subjecting the protein A eluted BC4 to hydroxylapatite chromatography as shown in Figure 2B.

Mentions: (A) Western blot analysis of BC2 (i), BC4 (ii) and ST2146 (iii) antibodies. A: pTn28; B: A–D fragment; C: tenascin. (B) Competitive ELISA. Biotinylated ST2146 was mixed with increasing concentrations of ST2146, BC4 or a not related IgG1 as competitors and plated on tenascin-coated plates. Binding was measured after addition of horseradish peroxidase–streptavidin and related chromogenic substrate.


Novel antitenascin antibody with increased tumour localisation for Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT).

De Santis R, Anastasi AM, D'Alessio V, Pelliccia A, Albertoni C, Rosi A, Leoni B, Lindstedt R, Petronzelli F, Dani M, Verdoliva A, Ippolito A, Campanile N, Manfredi V, Esposito A, Cassani G, Chinol M, Paganelli G, Carminati P - Br. J. Cancer (2003)

ELISA plates coated with tenascin at 5 μg ml−1 (A) or 0.5 μg ml−1 (B). BC4 peaks 1–3 were obtained by subjecting the protein A eluted BC4 to hydroxylapatite chromatography as shown in Figure 2B.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376359&req=5

fig4: ELISA plates coated with tenascin at 5 μg ml−1 (A) or 0.5 μg ml−1 (B). BC4 peaks 1–3 were obtained by subjecting the protein A eluted BC4 to hydroxylapatite chromatography as shown in Figure 2B.
Mentions: (A) Western blot analysis of BC2 (i), BC4 (ii) and ST2146 (iii) antibodies. A: pTn28; B: A–D fragment; C: tenascin. (B) Competitive ELISA. Biotinylated ST2146 was mixed with increasing concentrations of ST2146, BC4 or a not related IgG1 as competitors and plated on tenascin-coated plates. Binding was measured after addition of horseradish peroxidase–streptavidin and related chromogenic substrate.

Bottom Line: The hybridoma clone producing the monoclonal antitenascin antibody BC4, previously used for clinical applications, was found not suitable for further development because of the production of an additional, nonfunctional light chain.ST2146 was found able to bind human tenascin at an epitope strictly related, if not identical, to the antigenic epitope of BC4.It showed, compared to BC4, higher affinity and immunoreactivity and similar selectivity by immunohistochemistry.

View Article: PubMed Central - PubMed

Affiliation: Immunology Department, Sigma Tau SpA R&D, Pomezia, Rome, Italy. rita.desantis@sigma-tau.it

ABSTRACT
The Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT) method is based on intravenous, sequential administration of a biotinylated antibody, avidin/streptavidin and (90)Y-labelled biotin. The hybridoma clone producing the monoclonal antitenascin antibody BC4, previously used for clinical applications, was found not suitable for further development because of the production of an additional, nonfunctional light chain. In order to solve this problem, the new cST2146 hybridoma clone was generated. The monoclonal antibody ST2146, produced by this hybridoma, having the same specificity as BC4 but lacking the nonfunctional light chain, was characterised. ST2146 was found able to bind human tenascin at an epitope strictly related, if not identical, to the antigenic epitope of BC4. It showed, compared to BC4, higher affinity and immunoreactivity and similar selectivity by immunohistochemistry. Biodistribution studies of biotinylated ST2146 and three other monoclonal antitenascin antibodies showed for ST2146 the highest and more specific tumour localisation in HT29-grafted nude mice. On the overall, ST2146 appears to be a good alternative to BC4 for further clinical development of PAGRIT.

Show MeSH
Related in: MedlinePlus