Table I.

Characterization of hybridomas from ppc1-5H/Jκdel mice following PC-KLH immunization

HybridaIsotypetgbVHcMutationdAg-Binding Activitye
JHVHPC-HisPC-KLHssDNA
ppc1-5γ2b/λ1+7183.14300100100100
F2-6γ1/λ1+DQ52.2.4323333731
7183.1435 + nt del
F2-12γ1/λ1+DQ52.2.4322402916
7183.1436 + nt del
F1-5γ3/λ1SM7.4.63346194f120<5
F2-3γ1/λ1J558.1535814400<5
F2-4γ1/λ1J558.67.16646143374<5
F2-5γ1/λ1J558.83.189382283608
F1-6γ2b/λ1J558.6.963332913319
F2-2γ1/λ17183.2.33564917050
F1-3γ3/λ1Vgam3.82302614616
F1-8γ2b/λ17183-20.374527518<5
F2-9γ1/λ1J558.83.1893102421710
F2-8γ1/λ1VH37.13103283918
F1-7γ2a/λ17183.20.371172819<5
F1-4γ3/λ1J558.78.1821125218419
F1-9γ2b/λ17183.20.371132277<5
F2-1γ1/λ1VH61-1p332203428
F2-10γ1/λ17183.20.371121622<5
F2-11γ2a/λ1J558.83.1893831615<5
F2-7γ2a/λ17183.20.3711111119<5
Average mutational frequency5.3/VH3.3/Vλ
  • a Hybridomas were generated from splenic B cells of two ppc1-5H/Jκdel sd-tg mice following the third immunization with PC-KLH (see Materials and Methods).

  • b Presence or absence of ppc1-5H sd-tg was determined by PCR.

  • c VH and JH gene sequences were determined by RT-PCR sequencing. Individual genes were assigned based on the NCBI BLAST Ig Germline Gene Database. The 7183.20.37 and J558.83.189 VH genes were used by more than one hybridoma, but none of the hybridomas in these two VH gene groups were clonally related because they had different CDR3 junctions (data not shown).

  • d Number of mutations per V gene.

  • e Ag binding was determined by solid-phase ELISA as described in Materials and Methods. Relative binding activity was calculated based on OD at 2 h in ELISA and expressed as percentage of original ppc1-5 Ab OD. All samples were tested at the same time using the same concentration (2 μg/ml).

  • f Bold indicates binding avidity higher than ppcl-5 Ab.