Title :
Conjugation of Quantum Dots and JT95 IgM Monoclonal Antibody for Thyroid Carcinoma Without Abolishing the Specificity and Activity of the Antibody
Author :
Watanabe, Michiko ; Fujioka, Kouki ; Akiyama, Nobutake ; Takeyama, Hiroshi ; Manabe, Noriyoshi ; Yamamoto, Kenji ; Manome, Yoshinobu
Author_Institution :
Dept. of Mol. Cell Biol., Inst. of DNA Med., Tokyo, Japan
fDate :
3/1/2011 12:00:00 AM
Abstract :
Among the immunoglobulins, IgM class-antibodies are now considered to be potent immunological reagents for anticancer remedies. However, only a few reports are available about the effective labeling of IgM with enzymes, fluorescence, or other bioreactive reagents. Here, we report an effective application of luminescent semiconductive nanoparticles, quantum dots (QDs), as a labeling material of the IgM antibody. The CdSe carboxyl QDs were reacted with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysulfo- succinimide in 2-(morpholino) ethanesulfonic acid. The reacted QDs were then coupled to JT95 IgM antibody, which recognizes thyroid carcinoma associated antigen. The specificity and activity of the conjugates were tested by immunoblot, immunoquantitive assay and immunohistological imaging. The QDs were firmly conjugated with JT95 IgM monoclonal antibody. In immunoblot assay, QD-JT95 conjugates directly detected the target molecules without obstructing the binding site. In immunoquantitive assay, the conjugates could quantify the antigen in the range of 1.56-100 μg/mL. Also, QDs-labeled antibody detected the antigen on plasma membrane. Our results demonstrate that labeling of JT95 and other IgM class antibodies with QDs is feasible. This approach may be an important method for the medical application of IgM in the diagnosis and treatment of cancers.
Keywords :
II-VI semiconductors; biological organs; biomedical materials; biomedical optical imaging; cancer; luminescence; molecular biophysics; nanomedicine; nanoparticles; patient treatment; proteins; semiconductor quantum dots; 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride; 2-(morpholino)ethanesulfonic acid; CdSe carboxyl quantum dots; IgM class antibodies; IgM monoclonal antibody-quantum dot conjugation; JT95 IgM monoclonal antibody; N-hydroxysulfo-succinimide; antibody activity; antibody specificity; anticancer remedies; cancer diagnosis; cancer treatment; effective IgM labeling; immunoblot assay; immunoglobulins; immunohistological imaging; immunological reagents; immunoquantitive assay; luminescent semiconductive nanoparticles; plasma membrane antigen; thyroid carcinoma associated antigen; Biomembranes; Immune system; Labeling; Medical diagnostic imaging; Proteins; Quantum dots; Conjugation; IgM; quantum dot; thyroid carcinoma; Antibodies, Monoclonal; Antibody Specificity; Antigens; Cadmium Compounds; Cell Line, Tumor; Humans; Immunoglobulin M; Quantum Dots; Selenium Compounds; Spectrometry, Fluorescence; Thyroid Neoplasms;
Journal_Title :
NanoBioscience, IEEE Transactions on
DOI :
10.1109/TNB.2011.2125800