(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE (19) World Intellectual Property Organization Intemational Bureau (43) International Publication Date 24 March 2005 (24.03.2005) PCT ENT COOPERATION TREATY (PCT) (10) International Publication Number WO 2005/026191 A2 (51) International Patent Classification’ co7TK 21) International Application Number: PCT/KR2004/002383 (22) International Filing Da 17 September 2004 (17.09.2004) 25) Filing Language: English (26) Publication Language: English G0) Priority Data: PCT/KROSIO1913 18 September 2003 (18.09.2003) KR PCT/KRO302261 24 October 2003 (24.10.2003) KR 60/567, 844 3 May 2004 (03.05.2008) US (60/571,052 14 May 2004 (14.05.2008) US 1017601 12 August 2004 (12.08.2004) US ™ ™ (75) Applicants (forall designated States except US): POSCO IKR/KR]; 1 Goodong-dong, Nam-ku, Pohang: shi, uungsangbuk-do 790-300 (KR). POSTECH Founda- tion [KR/KRJ; San 31, Hyoje-dong, Nam-ku, Pohang-City, Kyungsangbuk-do 790-784 (KR). Inventors; and Inventors/Applicants for US only): PARK, Joon, Won IKR/KR]; 48-1303 Professor Apt, Jigok-dong, Namgu, Pohang, Gyeonsangbuk-do, 190-751 (KR). HONG, Bong, ‘Fin [KR/KR| #2-1005 Postech Graduate Apt. igok-dong, Namgu, Pohang, Gyeonsangbuk-do 790-751 (KR). CHOL, Young, Seo [KR/KR]; #2041704 Jugong New Town, 861, Jowor-dong, Jangan-gu, Suwon, Gyeonggi-do 440-200) (KR). OH, Soon, Jin [KR/KR]; #203-1405 Buyeong 2-cha Apt, Sadong, Gyeongsan, Gyeonsangbuk-do, 712-790 (KR). CHOL, Kwan, Yong [KR/KR]; #5-208 Mirane Api, San 89.3, Bongdeok-dong, Namgu, Daogu 705.020 (KR), [Continued on ness page] WO 2005/026191 A2 (S4) Title: SIZE-CONTROLLED MACROMOLECULE, (87) Abstract: The present application discloses a substrate that includes a molecular layer of regularly spaced size-contolled macromolecules comprising a polymer comprising brunched and linear regions in which 4 plurality of termini on the branched region are bound to the substrate, and a terminas of the linear region is FunctionalizedWO 2005/026191 A2 (74) Agents: YOU ME Patent & Law Firm; Seolim Bldg. (649-10 Yoksaa-dong, Kangnam-Ku, Seoul 135-080 (KR). (81) Designated States (unless otherwise indicated, for every (84) Designated States (unless otherwise indicated, for every Kind of regional protection available}: ARIPO (BW, GH, ‘TZ, UG, ZM, GM, KE, LS, MW, MZ, NA, TM, Eurasian (AM, SK, TR), OAPI (BF, BI, CF, C GW, ML, MR, NE, SN, TD, TO), Publis — without international search report and to be republished upon receipt of that report For two-leser codes and other abbreviations, refer to the "Guid- ‘nce Notes on Codes and Abbreviations” appearing at the begin hing ofeach regular issue of the PCT Gacete10 6 20 30 WO 2005/026191 PCT/KR2004/002383 Size-Controlled Macromolecule BACKGROUND OF THE INVENTION a) Field of the Invention The present invention relates to the field of hyperbranched macromolecules. The present invention relates to the field of functionalized substrates on which is bound the macromolecules. The present invention also relates to the field of functionalized size-controlled dendrimers and dendrons that are used to bind to a functionalized substrate at one end of the dendron and to a target-specific ligand on the other end. The present invention also relates to the field of combinatorial chemistry, specific protein detection methods, specific nucleic acid or nucleic acid/peptide hybrid detection methods using a functionalized substrate to which is bound a hyperbranched polymer linked to a probe biomolecule. b) Description of the Related Art Since the first report (Fodor et al., Nature 364, 555-556 (1993); Saiki et al., Proc. Natl. Acad. Sci. USA 86, 6230-6234 (1986)), DNA microarrays have attracted a great deal of attention because they allow high-throughput analysis of the DNA. sequence, genetic variations, and gene expression. It is known that this methodology requires improvement in terms of fidelity, reproducibility, and spot homogeneity that are essential for the standardization and application to human gene diagnosis (Hackett et al., Nature Biotechnology 21, 742-743 (2003). These shortcomings are caused mainly by the variations in the nature of the surface and molecular interlayer structures that are far from ideal. Likewise, the field of high- throughput target detection systems encompasses bioassays utilizing immobilized bioactive molecules and biomolecules. Here we show that DNA microarrays fabricated on a nanoscale-controlled surface discriminates single mismatched pairs as effectively as DNA does in solution. This approach provides an ideal DNA-microarray in which each probe DNA strand is given ample space enough to interact with an incoming target DNA with minimal steric hindrance. The dramatically increased discrimination efficiency promises the very reliable diagnosis of human genes. Moreover, the approach is general enough to be applied to various bioassays utilizing immobilized bioactive molecules and biomolecules.