Development of antibody functionalized magnetic nanoparticles for the immunoassay of carcinoembryonic antigen: a feasibility study for clinical use

Journal of Nanobiotechnology

Kai-Wen Huang²  Bing-Hsien Liu¹  Jui-Feng Chang¹  Chia-Shin Ho¹  Shieh-Yueh Yang¹  Che-Chuan Yang¹ 

[1]MagQu Co., Ltd., Sindian Dist, New Taipei City 231, Taiwan

[2]Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei 100, Taiwan

关键词: Colorectal cancer;    Assay;    Carcinoembryonic antigen;    Bio-magnetic nanoparticles;   

Others  :  1139420
DOI  :  10.1186/s12951-014-0044-6

 received in 2014-08-10, accepted in 2014-10-13,  发布年份 2014

【 摘 要 】

Background

Magnetic nanoparticles functionalized antibodies are used for in-vitro assays on bio-markers. This work demonstrates the synthesis of high-quality magnetic nanoparticles coated with antibodies against carcinoembryonic antigen (CEA). Various characterizations, such as particle size, particle suspension, bio-activity and the stability of bio-magnetic nanoparticles suspended in liquid, are studied. The properties for the assay of CEA molecules in serum are also studied. The assay method used is so-called immunomagnetic reduction.

Results

The results show that the effects of common materials in serum that interfere with detected signals are not significant. The low-detection limit is 0.21 ng/ml, which is well below the clinical threshold of 2.5 ng/ml.

Conclusions

The dynamic range for the assay of CEA molecules in serum is 500 ng/ml. By assaying serum CEA molecules from 24 normal controls and 30 colorectal-cancer patients, the threshold for the serum-CEA concentration to diagnose colorectal cancer is 4.05 ng/ml, which results in a clinical sensitivity of 0.90 and specificity of 0.87.

【 授权许可】

   
2014 Yang et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Hultman T, Ståhl S, Homes E, Uhlén M: Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support. Nucleic Acids Res 1989, 17:4937-4946.
• [2]Kondo A, Kamura H, Higashitani K: Development and application of thermo-sensitive magnetic immunomicrospheres for antibody purification. Appl Microbiol Biotechnol 1994, 41:99-105.
• [3]Franzreb M, Siemann-Herzberg M, Hobley TJ, Thomas ORT: Protein purification using magnetic adsorbent particles. Appl Microbiol Biotechnol 2006, 70:505-516.
• [4]Gånshirt-Ahlert D, Börjesson-Stoll R, Burschyk M, Garritsen HSP, Helmer E, Miny P, Velasco M, Walde C, Patterson D, Teng N, Bhat NM, Bieber MM, Holzgreve W: Detection of fetal trisomies 21 and 18 from maternal blood using triple gradient and magnetic cell sorting. Am J Reprod Immunol 1993, 30:194-201.
• [5]Pamme N, Wilhelm C: Continuous sorting of magnetic cells via on-chip free-flow magnetophoresis. Lab Chip 2006, 6:974-980.
• [6]Adamsa JD, Kim U, Soh HT: Multitarget magnetic activated cell sorter. Proc Natl Acad Sci U S A 2008, 105:18165-18170.
• [7]Kim DK, Zhang Y, Kehr J, Klason T, Bjelke B, Muhammed M: Characterization and MRI study of surfactant-coated superparamagnetic nanoparticles administered into the rat brain. J Magn Magn Mater 2001, 225:256-261.
• [8]Huang KW, Chieh JJ, Lin IT, Horng HE, Yang HC, Hong CY: Anti-CEA-functionalized superparamagnetic iron oxide nanoparticles for examining colorectal tumors in vivo. Nanoscale Res Lett 2013, 8:413-416. BioMed Central Full Text
• [9]Dobson J: Magnetic nanoparticles for drug delivery. Drug Dev Res 2006, 67:55-60.
• [10]Hergt R, Dutz S, Müller R, Zeisberger M: Magnetic particle hyperthermia: nanoparticle magnetism and materials development for cancer therapy. J Phys Condens Matter 2006, 18:S2919-S2934.
• [11]Kotitz R, Matz H, Trahms L, Koch H, Weitschies W, Rheinlander T, Semmler W, Bunte T: SQUID based remanence measurements for immunoassays. IEEE Trans Appl Supercond 1997, 7:3678-3681.
• [12]Enpuku K, Minotani T, Gima T, Kuroki Y, Itoh Y, Yamashita M, Katakura Y, Kuhara S: Detection of magnetic nanoparticles with superconducting quantum interference device (SQUID) magnetometer and application to immunoassays. Jpn J Appl Phys 1999, 38:L1102-L1105.
• [13]Hong CY, Wu CC, Chiu YC, Yang SY, Horng HE, Yang HC: Magnetic susceptibility reduction method for magnetically labeled immunoassay. Appl Phys Lett 2006, 88:1-3. 212512
• [14]Baselt DR, Lee GU, Natesan M, Metzger SW, Sheehan PE, Colton RJ: A biosensor based on magnetoresistance technology. Biosens Bioelectron 1998, 13:731-739.
• [15]Krause HJ, Wolters N, Zhang Y, Offenhäusser A, Miethe P, Meyer MHF: Magnetic particle detection by frequency mixing for immunoassay applications. J Magn Magn Mater 2007, 311:436-444.
• [16]Horng HE, Yang SY, Huang YW, Jiang WQ, Hong CY, Yang HC: Nanomagnetic particles for SQUID-based magnetically labeled immunoassay. IEEE Trans Appl Supercond 2005, 15:668-671.
• [17]Liao SH, Yang HC, Horng HE, Chieh JJ, Chen KL, Chen HH, Chen JY, Liu CI, Liu CW, Wang LM: Time-dependent phase lag of biofunctionalized magnetic nanoparticles conjugated with biotargets studied with alternating current magnetic susceptometor for liquid phase immunoassays. Appl Phys Lett 2013, 103:1-3. 243703
• [18]Shao H, Yoon TJ, Liong M, Weissleder R, Lee H: Magnetic nanoparticles for biomedical NMR-based diagnostics. Beilstein J Nanotechnol 2010, 1:142-154.
• [19][http://www.magnabiosciences.com/] webcite MagnaBioSciences, LLC.
• [20][http://www.magnisense.com/] webcite Magnisense Co., Ltd.
• [21][http://www.lifeassays.com/] webcite Lifeassays.
• [22][http://www.goldmag.com.cn/] webcite Xi'an Goldmag Nanobiotech Co., Ltd.
• [23][http://www.magqu.com/] webcite MagQu Co., Ltd.
• [24]Yang SY, Wang WC, Lan CB, Chen CH, Chieh JJ, Horng HE, Hong CH, Yang HC, Tsai CP, Yang CY, Cheng IC, Chung WC: Magnetically enhanced high-specificity virus detection using bio-activated magnetic nanoparticles with antibodies as labeling markers. J Virol Methods 2010, 164:14-18.
• [25]Jiang WQ, Yang HC, Yang SY, Horng HE, Hung JC, Chen YC, Hong CY: Preparation and properties of superparamagnetic nanoparticles with narrow size distribution and biocompatible. J Magn Magn Mater 2004, 283:210-214.
• [26]Chieh JJ, Yang SY, Horng HE, Yu CY, Lee CL, Wu HL, Hong CY, Yang HC: Immunomagnetic reduction assay using high-Tc superconducting-quantum-interference-device-based magnetosusceptometry. J Appl Phys 2010, 107:1-5. 074903
• [27]Yang CC, Yang SY, Chen HH, Weng WL, Horng HE, Chieh JJ, Hong CY, Yang HC: Effect of molecule-particle binding on the reduction in the mixed-frequency ac magnetic susceptibility of magnetic bio-reagents. J Appl Phys 2012, 112:1-4. 24704
• [28]Yang SY, Chieh JJ, Huang KW, Yang CC, Chen TC, Ho CS, Chang SF, Chen HH, Horng HE, Hong CY, Yang HC: Molecule-assisted nanoparticle clustering effect in immunomagnetic reduction assay. J Appl Phys Lett 2013, 113:1-5. 144903