Method for monitoring efficacy of a cancer therapy using circulating tumor cells as a biomarker
A tumor cell and therapy technology, used in radiotherapy, biomaterial analysis, X-ray/γ-ray/particle irradiation therapy, etc., which can solve the problems of low capture purity, hindering post-capture analysis of CTCs, and low sensitivity.
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Embodiment 1
[0034] Example 1: Materials and methods
[0035] Materials. Anti-human epithelial cell adhesion molecule (EpCAM) / TROP1 antibody (aEpCAM), anti-human epidermal growth factor receptor-2 (HER-2) / TROP1 antibody (aHER-2) and recombinant human E-selectin (E- Selectin) was purchased from R&D systems (Minneapolis, MN). Anti-human epidermal growth factor receptor (EGFR) antibody (aEGFR, N-20) was obtained from Santa Cruz Biotech (Dallas, TX). Epoxy functionalized glass surface Available from TeleChem International, Inc. (Sunnyvale, CA). PAMAM dendrimers (generation 7), bovine serum albumin (BSA), and all other chemicals were obtained from Sigma-Aldrich (St. Louis, MO) unless otherwise indicated, and unless otherwise indicated, Otherwise all were used directly without further purification.
[0036] Surface functionalization by immobilization of capture agents. Surface functionalization was performed using established methods (Myung et al., (2014) Anal. Chem.86(12):6088-94; Myung e...
Embodiment 2
[0043] Example 2: Surface preparation and UICHIP TM manufacture
[0044] UICHIP integrated with G7PAMAM dendrimer, E-selectin and antibody cocktail TM It was fabricated using previously described surface chemistry methods (Myung et al., (2011) Angew Chem. Int. Ed. Engl. 50(49):11769-72). Briefly, partially carboxylated G7PAMAM dendrimers were passed through heterobifunctional polyethylene glycol (PEG, COOH-PEG-NH 2 ) linker using amine coupling chemistry based on 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide / N-hydroxysulfosuccinimide (EDC / NHS) (Myung etc., (2011) Angew Chem.Int.Ed.Engl.50(49):11769-72) immobilized on the glass slide of epoxy resin functionalization. Then the antibody mixture (ABmix) of aEpCAM, aHER-2 and aEGFR was coupled by EDC / NHS (Myung et al., (2011) Angew Chem.Int.Ed.Engl.50(49):11769-72; Myung et al., ( 2014) Anal.Chem.86(12):6088-94), coupled to the carboxyl terminus of G7PAMAM dendrimers. Since this step allows the consumption of most of the prima...
Embodiment 3
[0045] Example 3: Patient Demographic Data
[0046] Patients with histologically confirmed primary cancers who underwent RT for tumor management were enrolled in this study. A total of 21 patients with rectal cancer (n=1), cervical cancer (n=1), prostate cancer (n=1), oral cavity cancer (n=2), sinus cancer ( n=3) or oropharyngeal cancer (n=13). Patient demographics and clinical information are summarized in Table 1. A baseline blood sample (pre-RT) was collected within one week of starting RT, usually on the day of the CT simulation used for RT planning or the day of pre-treatment patient preparation. During RT, samples were collected at up to 3 time points, including during the first week of RT (1W-RT), halfway through RT (in RT), and during the last week of RT (end of RT). Final samples were collected at least 4 weeks later than the last week of RT (post-RT).
[0047] Table 1
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[0049]
[0050] A total of 19 (90%) of 21 recruited patients had at least one ...
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