TABLE 3.
Titers and heparin binding properties of mutants
| Mutant virusa | Particle titer ofb:
|
Infectious titer (IU/ml)c | Particle-to-infectivity ratiod | Heparin bindinge | Empty/full ratiog | |
|---|---|---|---|---|---|---|
| A20/ml | Genome/ml | |||||
| rAAV2 (WT) | 1.5 × 1012 | 4.6 × 1011 | 1.8 × 1010 | 25 | + | 3.4 |
| H358A | <1.0 × 108 | <1.0 × 106 | <1.0 × 104 | N/Df | N/D | N/D |
| R447A | 1.2 × 1012 | 3.4 × 1010 | 1.3 × 109 | 25 | + | 35.9 |
| R459A | 9.1 × 1010 | 7.2 × 108 | <1.0 × 104 | >72,500 | + | 126.3 |
| R484A | 1.5 × 1011 | 3.0 × 1010 | <1.0 × 104 | >2,976,667 | +/− | 5.1 |
| R487A | 5.4 × 1011 | 2.2 × 1011 | 2.3 × 108 | 954 | +/− | 2.5 |
| H509A | 4.6 × 1010 | 2.3 × 109 | 6.9 × 105 | 3,285 | + | 20.3 |
| R513A | 2.9 × 1011 | 1.7 × 1010 | 1.6 × 108 | 106 | + | 17.9 |
| K532A | 1.1 × 1011 | 3.6 × 1010 | <1.0 × 104 | >3,633,333 | +/− | 3.0 |
| K544A | 2.0 × 1011 | 1.7 × 1010 | 8.3 × 108 | 20 | + | 11.9 |
| R566A | 5.1 × 1011 | 1.6 × 1010 | 7.4 × 108 | 21 | + | 32.6 |
| R585A | 5.0 × 1011 | 4.8 × 1010 | 1.7 × 107 | 2,812 | − | 1.4 |
| N587A | 4.4 × 1011 | 1.3 × 1010 | 7.7 × 107 | 165 | + | 34.7 |
| R588A | 2.4 × 1011 | 5.6 × 1010 | 3.0 × 106 | 18,521 | − | 4.2 |
| H526A, K527A | 1.4 × 1011 | 8.2 × 1010 | 5.5 × 107 | 1,489 | + | 1.8 |
| R585A, R588A | 1.2 × 1012 | 9.2 × 1011 | 1.9 × 107 | 48,421 | − | 1.2 |
| R585K | 1.3 × 1012 | 3.7 × 1010 | 4.0 × 108 | 92 | + | 35.4 |
| R585K, R588K | 1.4 × 1012 | 3.9 × 1010 | 8.9 × 107 | 436 | + | 34.9 |
| AAV1 | N/D | 3.7 × 1010 | 1.1 × 109 | 37 | +/− | N/D |
| AAV5 | N/D | 3.4 × 1010 | 3.2 × 106 | 10,692 | − | N/D |
| AAV5-HS | N/D | 8.0 × 108 | <1.0 × 104 | >80,000 | + | N/D |
Two letters flanking a number designate each mutant. The first letter is the one-letter abbreviation for the wild-type amino acid followed by its numerical position in VP1 followed by the one-letter abbreviation for the amino acid to which it was mutated.
A20 particle titers were determined as described in Materials and Methods using the A20 ELISA. Genomic titers were determined by RT-PCR.
Infectious titers were determined by GCA as described by counting GFP fluorescent cells.
Particle-to-infectivity ratio was calculated by dividing the average genomic titer as determined by RT-PCR by the average GCA titer.
Determined by heparin-agarose binding assay. +, >95% virus recovered in the eluate; +/−, >50% recovered in the eluate; −, <5% of virus recovered in the eluate.
N/D, not determined.
Empty-to-full ratio was determined by dividing the A20 particle titer by the average genomic titer.