101 



ADGG-CON A Conjugates 



Synthesis 



14 

 The results of the ' C-hippuric acid studies indicated 



that reaction at pH 5 in phosphate buffer with relatively 



low concentrations of protein and carbodiimide would result 



in formation of Con A conjugates with a defined number of 



substitutions and minimal perturbation of the native lectin 



structure. These conditions were used for the synthesis 



of H-DM-ADGG-Con A conjugates in order to verify the 



reaction conditions and to produce a labeled ADGG-Con A 



conjugate for cell binding experiments. 



3 

 After completion of the synthesis, the H-DM-ADGG-Con A 



containing reaction mixture was applied to a Sephadex G-75 

 column and isolated by absorption to the gel. Figure 21 

 shows the chromatographic profile as determined by total 

 radioactivity eluted and TCA precipitable acitvity. The 

 majority of the radioactivity emerged from the column as a 

 nonprecipitable fraction in a single peak. Following addi- 

 tion of . 1M D-glucose to the eluant, H-DM-ADGG-Con A con- 

 jugate was eluted in a single peak that was 100% precipitable 

 by TCA. This peak contained greater than 90% of the total 

 lectin present in the reaction. Similar reaction conditions 

 were used to produce other conjugates of ADGG and Con A that 

 were used for examining the interaction of ADGG-Con A conju- 

 gates with cultured cells. The ADGG-Con A conjugates were 

 isolated by absorption chromatography to Sephadex G-75. 



