SBIR Phase II: Novel Chemical Methods for Labeling Small Peptides

这个小企业创新研究二期项目将进一步开发使用化学发光检测方法替代小肽免疫测定的放射性方法。生物样品中小肽的浓度通常用生物测定法或放射免疫测定法测定。生物测定往往是不准确和繁琐的执行，而放射免疫测定使用'251标记肽作为检测方法。使用i25I进行这些检测的成本、处理、环境、稳定性和灵敏度问题使替代检测系统具有吸引力。在第一阶段，我们证明了化学发光可以用于测量小肽，加压素，速度和灵敏度。我们成功地制造出稳定的抗利尿激素缀合物，其速度和灵敏度都有所提高。在第一阶段展示的化学方法必须进一步发展和完善。肽类似物用于提高抗血清和偶联到化学发光检测系统的用途需要进一步测试。我们能够使用抗利尿激素的类似物，合成在实验中具有更好的结合和灵敏度的缀合物。我们希望探索利用这些和类似的肽类似物提高抗血清免疫原的可能性。我们相信使用这些类似物，其中的偶联位置已被选择从天然肽中化学分离，可能产生具有更好的特异性和反应性的抗血清。Assay designs是一家在开发、营销和销售使用化学发光的新型免疫分析测试试剂盒方面拥有成熟记录的公司。我们定期在国家科学杂志上刊登广告，并在日本、奥地利、德国和瑞士设有经销商。化学发光技术应用于测量分析，其中唯一敏感的，可重复的方法是放射性的，具有巨大的潜力。对HI的使用和处理的关注使得化学荧光法可以取代RIA系统，提高这些检测的灵敏度和速度。一个RLA制造商有139个不同的套件，每个套件售价高达450美元。具有速度和灵敏度优势的非放射性试剂盒的市场是巨大的

DMI-9531248 Hart This Small Business Innovation Research Phase II project will further develop ways to replace radioactivity for small peptide immunoassays using chemiluminescence detection methods. The concentration of small peptides in biological samples are normally measured using bioassays or by radioimmunoassay methods. Bioassays tend to be inaccurate and cumbersome to perform, while radioimmunoassays use '251 labeled peptides as a detection method. The cost, handling, environmental, stability and sensitivity concerns of using i25I for these assays makes alternative detection systems attractive. In Phase I we demonstrated that chemiluminescence can be applied to measure a small peptide, vasopressin, with speed and sensitivity. We managed to make stable conjugates of vasopressin that demonstrated increased speed and sensitivity. The chemical methods demonstrated in Phase I have to be further developed and refined. The use of peptide analogs for use in raising antisera and also for conjugation to chemiluminescent detection systems need to be further tested. We were able, using analogs of vasopressin, to synthesize conjugates that have better binding and sensitivity in the assay. We wish to explore the possibility of raising antisera to immunogens using these and similar peptide analogs. We believe that the use of these analogs, where the conjugation position has been chosen to be chemically isolated from the native peptide, may yield antisera with better specificity and reactivity. Assay designs is a company with a proven, track record of developing, marketing and selling novel immunoassay test kits using chemiluminescence. We advertise in national scientific magazines on a regular basis and have distributors in Japan and in Austria, Germany and Switzerland. The applications of chemiluminescence technology to the measurement of analyses where the only sensitive, reproducible methods are radioactive have tremendous potential. The concerns with the use and disposal of HI make chemilumi nescence the method that can replace this RIA system and improve the sensitivity and speed of these assays. One RLA manufacturer has 139 different kits selling for up to $450 per kit. The market for non-radioactive kits with the advantages of speed and sensitivity is substantial