Hexa-histidine tag as a novel alternative for one-step direct labelling of a single-chain Fv antibody fragment with 99m Tc.

From genetic material of hybridoma cells, we have generated a recombinant single-chain antibody fragment (scFv antibody) specific to carcinoembryonic antigen (CEA), which can substitute an intact murine monoclonal immunoglobulin G1 (IgG1) antibody, also developed by our group, and used in clinical practice for many years. In this paper, we examine a novel one-step method for direct 99mTc labelling of a recombinant anti-CEA scFv fragment through a C-terminal peptide tag containing a six-histidine sequence. This C-terminal peptide tag does not affect antigen binding, and was employed as a strategy for the one-step method of direct 99mTc labelling of a recombinant antibody fragment, based on the criteria of Zamora and Rhodes (Zamora PO, Rhodes BA. Imidazoles as well as thiolates in proteins bind technetium-99m. Bioconj Chem 1992; 3: 493-498). This is a novel technique for the rapid labelling of molecules, suitable for in vivo trials. The method yields >95% labelling efficiency without major effects on biological or in vitro stability.

[Radioimmunolocalization of head and neck tumors with a new monoclonal antibody against the EGF receptor. Preliminary results]. / Radioinmunolocalización de tumores de cabeza y cuello con un nuevo anticuerpo monoclonal contra el receptor del EGF. Resultados preliminares.

OBJECTIVE: To show the results of Radioimmunodetection (RID) using Cuban monoclonal antibody (MoA) anti-egf-ior-egf-r3 labeled with 99mTc for the detection of primary tumors and/or metastases of head and neck epithelial malignant tumors. MATERIAL AND METHODS: Thirteen patients aged from 16-62 years (mean: 54.8 years) with primary tumors and metastases in the regional cervical lymph nodes were retrospectively evaluated. The labelling dose was 1480-2220 MBq (40-60 mCi). Planar images were performed after 10 minutes and together with SPECT images after 18-24 hours following MoA administration. The matrix was 128 x 128. SPECT images were reconstructed using the Butterworth 4/16 filter. A positive result was defined when the lesions were visualized. RESULTS: RID localized 11 of the known primary tumors in the 13 patients studied. In the two other patients, in whom the primary tumor site was unknown, cervical lymph node metastases were found. The results of 3 of the RID were false negative but in the other 10, RID was able to localize the primary tumor and cervical lymph node metastases. Sensitivity was 77 % and the predictive positive value was 100 %. CONCLUSIONS: The results of this study show that the Cuban MoA ior-egf-r3 can be employed for RID of primary head and neck tumors and their metastases. The radiation dose is adequate for RID.

[Radioimmunoscintigraphy of colorectal cancer using the anti-CEA monoclonal antibody BW 431/26. Final results]. / Radioinmunocentellografía del cáncer colorrectal empleando el anticuerpo monoclonal BW 431/26 anti-CEA. Resultados finales.

The anti-carcinoembryonic antigen (CEA) antibody, BW 431/26 (Scintimun CEA, Behringwerke, Marburg, Germany ) labeled with technetium pertechnectate (99mTc), is an intact immunoglobulin G1, monoclonal antibody that has been used to image colorectal cancer. Planar and SPECT images of chest, abdomen and pelvis were performed at 10 minutes, 4-6 and 18-24 hours after the intravenous antibody injection. 44 patients were studied and the pathological antibody concentration localization by radioimmunoimaging (RI) were correlated with surgical, clinical and other imaging modality findings to validate the RI. The RI was positive in 29 patients and negative in the other 15 patients. The CEA and CA 19.9 were elevated in the serum of some patients with primary tumors or recurrence. The HAMA were determined in all the patients before and after the RI.