Skip to main content
. Author manuscript; available in PMC: 2014 Aug 1.
Published in final edited form as: J Immunol. 2013 Jul 3;191(3):1260–1275. doi: 10.4049/jimmunol.1300770

Table 3. Individual Ag reactivities and Ig usage properties of hybridomas derived from 2F5 and 4E10 complete KI splenic B cells.

Reactivity Profilea
 Ig Usage Profilesb
Clone ID No. MPER CL Neutralization
Ability		 VH
Rearrangeemnt
Used		 VH Mutation
Type/Location		 VL
Rearrangemen
t Used		 VL Mutation
Type/Location
4E10-V2-3 CL1 +++ N/A +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-5 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-6 CL4 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-10 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-12 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-13 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-20 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-30 CL1 +++ N/A +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-31 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-34 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-35 CL4 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-41 CL3 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-47 CL5 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-56 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-60 CL5 +++ +++ +++ KI 4E10Vh VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-62 CL1-1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-85 CL1 +++ N/A +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-98 CL1-1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-100 CL2 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-101 CL2 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-104 CL2 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-105 CL1 +++ N/A +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-108 CL8 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-112 CL7 +++ N/A N/A KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-114 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-122 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-123 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-126 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-127 CL4 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-131 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-134 CL3 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-141 CL2-1 +++ N/A +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V2-143 CL3-1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-1 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-3 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-6 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-10 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-16 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-17 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-18 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-19 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-22 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-24 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-27 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-33 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-35 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-36 CL1 +++ +++ +++ KI 4E10VH VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-37 CL1 +++ +++ +++ KI 4E10Vh VDJ no mutation KI 4E10VL VDJ no mutation
4E10-V3-40 CL1 +++ +++ +++ KI 4E10VH VDJ
J558.26.116/DFL		 no mutation KI 4E10VL VDJ no mutation
4E10-V2-49 CL3 - - - 16.1/partial
4E10VHc
J558.50.143/DSP		 N/A KI 4E10VL VDJ no mutation
4E10-V2-50 CL3 - - - 2.2/ partial
4E10VH 		N/A KI 4E10VL VDJ no mutation
2F5-V101-1 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-9 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-11 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-14 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-29 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-30 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-33 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-37 CL1 +++ ++ +++ KI 2F5Vh VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-50 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-74 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-77 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-82 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-103 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-115 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation N/A N/A
2F5-V101-124 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation N/A N/A
2F5-V101-125 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-130 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation N/A N/A
2F5-V101-136 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-143 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation N/A N/A
2F5-V101-160 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-165 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-175 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-202 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-215 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-247 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation N/A N/A
2F5-V101-256 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-276 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-282 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-299 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
2F5-V101-305 CL1 +++ ++ +++ KI 2F5VH VDJ no mutation KI 2F5VL VDJ no mutation
a

MPER, CL reactivities and neutralization ability were determined as described in Materials and Method. Criteria for positivity were arbitrarily set at saturating concentrations, and relative to control mAbs as follows: (+++) >80% of (4E10-V2-6 CL4 or V3-1.4) binding, (++) = 50-80% of (4E10-V2-6 CL4 or V3-1.4) binding, and (+) = 25-50% of (4E10-V2-6 CL4 or V3- 1.4) binding, but >13H11 (or AID 3G11) binding.

b

cDNAs from cloned hybridoma lines were amplified using leader-specific and K-specific primers, and PCR products were cloned and sequenced in both orientations as described in the Materials and Methods. KI VH and VL regions were analyzed for mutations using Lasergene software.

c

Heavy chains that cannot align to 4E10 VH were analyzed using the Ig BLAST algorithm. These 2 clones keep 3′end partial 4E10 VH sequence but replace their 5’end with endogenous VH segments in a region of the 4E10 VH that coinciding with a cryptic Recombination Signal Sequence (RSS) containing a perfect consensus embedded nonamer and heptamer (70).

d

N/A: not available.