Loftager M K, Koch C, Hellung-Larsen P, Andersen V
J Immunol Methods. 1987 Aug 24;102(1):65-9. doi: 10.1016/s0022-1759(87)80010-8.
DNA was conjugated to sheep red blood cells (SRBC) by chemical methods using CrCl3, poly-L-lysine or methylated bovine serum albumin as conjugation agents and by a physical method where conjugation was accomplished by incubation at 45 degrees C. The degree of conjugation was estimated using 32P-DNA (mean size 1 kbase pairs). Employing the CrCl3 method 5.8 +/- 3.6 micrograms DNA were conjugated per 10(8) SRBC at a concentration of 70 micrograms DNA/10(8) cells. At the same DNA concentration in the incubation medium 3.0 +/- 0.6 microgram DNA/10(8) cells were conjugated by poly-L-lysine, 4.1 +/- 0.8 microgram DNA/10(8) cells by methylated bovine serum albumin and approximately 4 micrograms DNA/10(8) cells when the cells were incubated at 45 degrees C. Cells conjugated with DNA by CrCl3 showed linearly increasing conjugation with increasing concentration of DNA. Cells conjugated by poly-L-lysine (pLL) or methylated bovine serum albumin seemed to be saturated by DNA at 30 micrograms DNA per 10(8) cells. At 45 degrees C the spontaneous adhesion of DNA to SRBC increased in the concentration range investigated. The degree of conjugation of DNA to SRBC was influenced by pH, and Ca2+.pLL-conjugated DNA-SRBC, but none of the other preparations were lysed in a hemolytic assay using anti-DNA antiserum from a patient with systemic lupus erythematosus.
采用化学方法，以三氯化铬、聚-L-赖氨酸或甲基化牛血清白蛋白作为偶联剂，将DNA与绵羊红细胞（SRBC）偶联；还采用了物理方法，即通过在45℃孵育来完成偶联。使用32P-DNA（平均大小为1千碱基对）估计偶联程度。采用三氯化铬方法，每10^8个SRBC在DNA浓度为70微克/10^8个细胞时偶联5.8±3.6微克DNA。在孵育培养基中相同DNA浓度下，聚-L-赖氨酸可使每10^8个细胞偶联3.0±0.6微克DNA，甲基化牛血清白蛋白可使每10^8个细胞偶联4.1±0.8微克DNA，当细胞在45℃孵育时，每10^8个细胞约偶联4微克DNA。用三氯化铬偶联DNA的细胞，其偶联程度随DNA浓度增加呈线性增加。用聚-L-赖氨酸（pLL）或甲基化牛血清白蛋白偶联的细胞，当DNA浓度达到每10^8个细胞30微克时似乎已饱和。在45℃时，在所研究的浓度范围内，DNA与SRBC的自发黏附增加。DNA与SRBC的偶联程度受pH值和Ca2+影响。pLL偶联的DNA-SRBC，但其他制剂在使用系统性红斑狼疮患者的抗DNA抗血清进行的溶血试验中均未裂解。
