SBIR Phase II: Novel Chemical Methods for Labeling Small Peptides

DMI-9531248 HART这个小型企业创新研究第二阶段项目将进一步开发使用化学发光检测方法取代放射性小肽免疫分析的方法。通常使用生物测定法或放射免疫测定法来测量生物样品中小肽的浓度。生物测定往往不准确且操作繁琐，而放射免疫分析使用‘251标记多肽作为检测方法。使用i25I进行这些检测的成本、操作、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法可以应用于快速和灵敏地测量一种小肽--加压素。我们成功地制造了稳定的加压素结合物，显示出更快的速度和更高的敏感性。第一阶段中展示的化学方法还需要进一步发展和完善。用于提高抗血清和与化学发光检测系统的偶联的多肽类似物的使用需要进一步测试。我们能够使用加压素的类似物来合成在检测中具有更好的结合力和灵敏度的结合物。我们希望探索使用这些和类似的多肽类似物来提高免疫原抗血清的可能性。我们认为，使用这些类似物，其中连接位置已被选择为与天然多肽化学分离，可能会产生更好的特异性和反应性的抗血清。分析设计公司是一家在开发、营销和销售使用化学发光技术的新型免疫分析测试试剂盒方面拥有经过验证的跟踪记录的公司。我们定期在国家科学杂志上做广告，并在日本、奥地利、德国和瑞士设有分销商。化学发光技术在分析测量中的应用具有巨大的潜力，在这些分析中，唯一灵敏、可重复的方法是放射性的。人们对HI的使用和处理的关注使化学发光检测成为可以取代这种RIA系统的方法，并提高了这些检测的灵敏度和速度。一家RLA制造商拥有139种不同的试剂盒，每个试剂盒的售价高达450美元。非放射性试剂盒具有速度快、灵敏度高等优点，市场前景广阔

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