Development of radioimmunoconjugate for diagnosis and management of head-and-neck subclinical cancer and colorectal carcinoma

ABSTRACT Scientific innovations in diagnostic methods are important drivers of cancer control and prevention. Noninvasive imaging of the epidermal growth factor receptor (EGFR) in head-and-neck squamous, cell carcinoma and colorectal cancer could be valuable to select patients for EGFR-targeted therapy, as well as to monitor the efficacy and occurrence of resistance to immunotherapy. In order to develop the first Brazilian radioimmunoconjugate for diagnosis, Cetuximab has been conjugated to p-SCN-Bn-DTPA chelator and radiolabeled with Indium-111. The conjugation methodology was optimized using different mAb:DTPA molar ratios, time was then reduced for immunoconjugate preparation, besides the protein recovery' percentage increased after purification (m = 83.8 ± 0.91 %). The stability of Cetuximab-DTPA at - 20 oC was evaluated for six months, and its integrity was greater than 90% (m =93.9 ± 1.5%, N = 24). The radioimmunoconjugate with specific activity of 185 MBq/mg showed radiochemical purity above 95% (m=96.8 ± 1.31 %, N = 15). We conclude that the radioimmunoconjugate 111In-DTPA-cetuximab is stable and may be applied to the diagnosis of EGFR-positive tumors.

Determinação dos fatores de recuperação do 111In e do 99mTc na quantificação de atividade com imagens SPECT / Determination of 111In and 99mTc recovery in the quantification of activity with SPECT imaging

OBJETIVO: Determinar, experimentalmente, os coeficientes de recuperação do 111In e do 99mTc usando imagens SPECT. MATERIAIS E MÉTODOS: Quatro diferentes concentrações de 111In e de 99mTc foram usadas para quantificar a atividade em esferas de diferentes tamanhos. As imagens foram obtidas com um equipamento híbrido SPECT/CT, com dois detectores. A reconstrução das imagens foi realizada usando o método iterativo ordered subset expectation maximization (OSEM). A correção de atenuação foi realizada com o uso de um mapa de atenuação e a correção de espalhamento foi realizada usando a técnica das janelas de energia. RESULTADOS: Os resultados mostraram que o efeito do volume parcial foi observado de forma mais significativa para as esferas com volume < 6 ml. Para o 111In, os resultados mostram uma dependência com relação às concentrações usadas nas esferas e ao nível de background usado. Para o 99mTc, pôde-se observar uma tendência à subestimação dos resultados quando os níveis mais altos de background foram utilizados. CONCLUSÃO: É necessário usar os fatores de correção para compensar o efeito do volume parcial em objetos com volume < 6 ml para ambos os radionuclídeos. A subtração das contagens espúrias presentes nas imagens SPECT foi o fator que mais influenciou na quantificação da atividade nessas esferas.

OBJECTIVE: To experimentally determine the 99mTc and 111In activity recovery coefficients in SPECT imaging. MATERIALS AND METHODS: Four different 99mTc and 111In concentrations were utilized for quantifying activity in spheres of four different sizes. Images were obtained with a hybrid dual-head SPECT-CT imaging system. The ordered subset expectation maximization (OSEM) iterative method was utilized for images reconstruction. An attenuation map was utilized for attenuation correction, and the multiple energy window technique for scattering correction. RESULTS: Results for spheres < 6 ml in volume were significantly affected by the partial volume effect. For 111In quantification, results show a dependence on sphere concentrations and background levels. For 99mTc quantification, there was a tendency towards values underestimation with higher background levels. CONCLUSION: Correction factors must be utilized for compensating the partial volume effect on objects with < 6 ml in volume for both radionuclides. Background subtraction to compensate spurious count present on SPECT images has a significant influence on the quantification of activity, especially for the smaller objects.

Biodistribution and Hepatic Metabolism of Galactosylated 111In-Antibody-Chelator Conjugates: Comparison with 111In-Antibody-Chelator Conjugates / 대한핵의학회잡지

PURPOSE: To evaluate the use of monoclonal antibody (MoAb) as a carrier of the receptor-binding ligand, the receptor mediated uptake into liver and subsequent metabolism of (111) In-labeled galactosylated MoAb-chelator conjugates were investigated and compared with those of (111) In labeled MoAb. MATERIALS AND METHODS: T101 MoAb, IgG2 against human lymphocytic leukemic cell, conjugated with cyclic DTPA dianhydride (DTPA) or 2-p-isothiocyanatobenzyl-6-methyl-DTPA (1B4M) was galactosylated with 2-imino-2-methoxyethyl-1-thio-beta-D-galactose and then radiolabeled with (111) In. Biodistribution and metabolism study was performed with two (111) In-conjugates in mice and rats. RESULTS: (111) In-labeled T101 and its galactosylated conjugates were taken to the liver by the time, mostly within 10 min. However DTPA conjugate was retained longer in the liver than the 1B4M conjugate (55% vs 20% of injected dose at 44 hr). During this time, the radiometabolite of DTPA conjugate was excreted similarly into urine (24%) and feces (17%). The radiometabolite of 1B4M was excreted primarily into feces (68%) rather than urine (8%). Size exclusion HPLC analysis of the bile and supernatant of liver homogenate showed two peaks, the first (35%) with the retention time (Rt) identical to IgG and the second (65%) with Rt similar to free 111In at 3 hr post-injection for the 1B4M conjugate, indicating that the metabolite is rapidly excreted through the biliary system. In contrast to DTPA conjugate, the small (111) In-DTPA-like metabolite was the major radioindium component (90%) in the liver homogenate as early as 3 hour post-injection, but the cumulative radioindium activity in feces was only 17% at 44 hour, indicating that the metabolite from DTPA conjugate does not clear readily through the biliary tract. CONCLUSION: The galactosylation of the MoAb conjugates resulted in higher hepatocyte uptake and enhanced metabolism, compared to those without galactosylation. Metabolism of the MoAb-conjugates is different between compounds radiolabled with different chelators due to different characteristics of radiometabolites generated in the liver.