军事医学
軍事醫學
군사의학
BULLETIN OF THE ACADEMY OF MILITARY MEDICAL SCIENCES
2014年
5期
341-346
,共6页
陈宇%杨静%李新颖%周婷婷%林周%罗龙龙%乔春霞%吕明%黎燕%沈倍奋%冯健男
陳宇%楊靜%李新穎%週婷婷%林週%囉龍龍%喬春霞%呂明%黎燕%瀋倍奮%馮健男
진우%양정%리신영%주정정%림주%라룡룡%교춘하%려명%려연%침배강%풍건남
抗体可变区%稳定性%可及性表面积%空间结构
抗體可變區%穩定性%可及性錶麵積%空間結構
항체가변구%은정성%가급성표면적%공간결구
antibody variable region%stability%accessible area%spatial structure
目的：通过研究抗体轻、重链间的结合能与其构象特征、理化性质间的内在关系，建立相应的数学模型，合理评价抗体分子的热稳定性，为抗体分子的设计、合理改造及亲和力成熟奠定基础。方法采用生物信息学和计算生物学相关方法对已有晶体结构的抗体结构信息进行分析，通过距离几何学、计算机图形学技术对抗体可变区的构象特征进行研究；基于分子间氢键形成理论、反应自由能理论，从热力学、动力学对抗体分子轻、重链可变区相互作用的动态结构及能量特征进行探讨；借助统计学原理、非线性拟合、回归分析等分析手段对抗体轻、重链作用能与其构象特征、理化性质建立相关性分析。结果通过分子模拟与统计学分析，抗体的轻、重链可变区结合能与其轻、重链间的氢键形成数目、范德华作用均存在线性相关性；与抗体理化性质存在基于多项式的非线性相关性。基于关键位置氨基酸的使用频率以及建立的分析模型，针对无法获得稳定工程细胞株的抗蓖麻毒素人源化功能抗体进行合理的评价及优化，借助理论指导，获得抗蓖麻毒素人源化抗体稳定的工程细胞株。结论抗体可变区的自身结构（构象、理化性质）对其稳定性有很大影响，可以通过抗体结构优化来提高抗体的稳定性。
目的：通過研究抗體輕、重鏈間的結閤能與其構象特徵、理化性質間的內在關繫，建立相應的數學模型，閤理評價抗體分子的熱穩定性，為抗體分子的設計、閤理改造及親和力成熟奠定基礎。方法採用生物信息學和計算生物學相關方法對已有晶體結構的抗體結構信息進行分析，通過距離幾何學、計算機圖形學技術對抗體可變區的構象特徵進行研究；基于分子間氫鍵形成理論、反應自由能理論，從熱力學、動力學對抗體分子輕、重鏈可變區相互作用的動態結構及能量特徵進行探討；藉助統計學原理、非線性擬閤、迴歸分析等分析手段對抗體輕、重鏈作用能與其構象特徵、理化性質建立相關性分析。結果通過分子模擬與統計學分析，抗體的輕、重鏈可變區結閤能與其輕、重鏈間的氫鍵形成數目、範德華作用均存在線性相關性；與抗體理化性質存在基于多項式的非線性相關性。基于關鍵位置氨基痠的使用頻率以及建立的分析模型，針對無法穫得穩定工程細胞株的抗蓖痳毒素人源化功能抗體進行閤理的評價及優化，藉助理論指導，穫得抗蓖痳毒素人源化抗體穩定的工程細胞株。結論抗體可變區的自身結構（構象、理化性質）對其穩定性有很大影響，可以通過抗體結構優化來提高抗體的穩定性。
목적：통과연구항체경、중련간적결합능여기구상특정、이화성질간적내재관계，건립상응적수학모형，합리평개항체분자적열은정성，위항체분자적설계、합리개조급친화력성숙전정기출。방법채용생물신식학화계산생물학상관방법대이유정체결구적항체결구신식진행분석，통과거리궤하학、계산궤도형학기술대항체가변구적구상특정진행연구；기우분자간경건형성이론、반응자유능이론，종열역학、동역학대항체분자경、중련가변구상호작용적동태결구급능량특정진행탐토；차조통계학원리、비선성의합、회귀분석등분석수단대항체경、중련작용능여기구상특정、이화성질건립상관성분석。결과통과분자모의여통계학분석，항체적경、중련가변구결합능여기경、중련간적경건형성수목、범덕화작용균존재선성상관성；여항체이화성질존재기우다항식적비선성상관성。기우관건위치안기산적사용빈솔이급건립적분석모형，침대무법획득은정공정세포주적항비마독소인원화공능항체진행합리적평개급우화，차조이론지도，획득항비마독소인원화항체은정적공정세포주。결론항체가변구적자신결구（구상、이화성질）대기은정성유흔대영향，가이통과항체결구우화래제고항체적은정성。
Objective To study the intrinsic relationships between the binding energy of the antibody light and heavy chains and the conformational characteristics , physical and chemical properties , and to establish a corresponding mathemat-ical model and evaluate the thermal stability of the antibody molecules , which contribute to the antibody design , optimiza-tion and affinity maturation .Methods Based on bioinformatics and computational biology methods , the antibody′s structur-al information with the crystal diffraction data was analyzed .The conformational character of the variable domain of the antibody was studied using distance geometry and computer graphics technology .With the aid of the intermolecular hydrogen bond formation theory and the reaction free energy theory , the dynamic structure and energy characteristics be-tween the heavy and light chain variable regions of the antibody were studied .Furthermore , using nonlinear fitting and regression analysis, a mathematical model was set up .Results According to simulation and statistic analysis , there was a linear relationship between the binding energy and the number of the intermolecular hydrogen bonding , Van der Waals interaction of the heavy and light chains of the antibody .There was polynomial correlation between the binding energy and the physicochemical properties of the antibody .Using the frequency of amino acid position and the established model , the humanized anti-ricin antibody , which could not obtain the stable engineering cell line , was evaluated and optimized .The stable engineering cell line of the humanized anti-ricin antibody was obtained in the experiment .Conclusion The self structure of the antibody variable region ( conformation and physicochemical properties ) has much effect on its stability . The antibody stability can be improved by structural optimization .
