An efficient system to generate monoclonal antibodies against membrane-associated proteins by immunisation with antigen-expressing mammalian cells

Table 1 Specificity of hybridoma culture supernatants

	IgG ELISA screen^a			IFA screen on transfected and non-transfected HEK cells^b
Antigen	total^c	IgG positive^d		total^e		negative^f		Pf protein specific^g		HEK cell specific^h		anchor specificⁱ
PFF0620c	1152	335	29%	142	42%	86	60%	25	18%	30	21%	1	0.7%
PFD1130w	1152	294	26%	294	100%	200	68%	10	3%	83	28%	1	0.3%
PF14_0325	1152	202	18%	202	100%	125	62%	2	1%	68	34%	7	3.0%

^a For each fusion hybridoma supernatants of 12 96-well-plates were analysed for the presence of mouse IgG by sandwich-ELISA
^b Wells screened positive by IgG ELISA were tested for binding to the corresponding transfected and the non-transfected HEK cells by IFA
^c Number of wells analysed by ELISA
^d Number and percentage of all tested wells that were IgG positive
^e Number and percentage of IgG positive wells that were analysed by IFA
^f Number and percentage of all wells tested by IFA that showed no reactivity in IFA
^g Number and percentage of all wells tested by IFA that only stained the transfectant cell line used for the immunisation of mice
^h Number and percentage of all wells tested by IFA that stained both transfected and non-transfected HEK cells by IFA
ⁱ Number and percentage of all wells tested by IFA that stained all three transfectants, but were negative for non-transfected HEK cells

ISSN: 1472-6750