In-house ELISA protocols for capsid p24 detection of diverse HIV isolates

Table 1 Variations between ELISA protocols

Features	ABR	ANG	SB	RND
Working volume	25 µL	25 µL	25 µL	50 µL
Working concentration of capture antibody	10 µg/mL	10 µg/mL	2 µg/mL	4 µg/mL
Washing buffer post-coating	TBS-based buffer	PBS-based buffer	PBS-based buffer	PBS-based buffer
Blocking step post-coating	No	Yes	Yes	Yes
Working concentration of detection antibody	60 ng/mL	60 ng/mL	100 ng/mL	125 ng/mL
Detection antibody	AP conjugated	AP conjugated	HRP conjugated	Unconjugated
Incubation period post-antibody addition	1 h	1 h	1 h	2 h
Washing buffer post-antibody addition	PBS-based buffer	PBS-based buffer	PBS-based buffer	PBS-based buffer
Additional conjugation	NA	NA	NA	HRP
Substrate reagent	LumiPhos Plus	LumiPhos Plus	LumiPhos-HRP	LumiPhos-HRP
Incubation period post-substrate addition	20 min	20 min	2 min	2 min
Lower range	0.014 ng/mL	0.17 ng/mL	0.018 ng/mL	0.015 ng/mL
Upper range	5 ng/mL	5 ng/mL	5 ng/mL	3 ng/mL

ISSN: 1743-422X